Risk Perceptions of Little Cigar and Cigarillo Smoking Among Adult Current Cigarette Smokers.
Few studies have examined the perceptions of risk of little cigar and cigarillo (LCC) smoking among cigarette smokers, which is important for expanding regulatory policies and developing prevention programs. We examined current cigarette smokers' perceived harm of LCC smoking, and determined whether these perceptions were associated with susceptibility and intention to continue smoking LCCs. Data were from the 2014 Tobacco Products and Risk Perceptions Survey of a probability sample of 5717 US adults. Data were analyzed for a subsample of 1191 current cigarette smokers who were stratified into three groups: (1) dual current cigarette smokers who had ever used LCCs, (2) current smokers susceptible to LCC smoking, and (3) current smokers who were not susceptible to LCC smoking. Overall, 47.2% of participants were dual smokers, 12.7% were susceptible to LCC smoking, and 40.1% were not susceptible. Perceptions of risk of LCCs varied across the groups. Dual smokers were more likely to perceive that daily LCC smoking is "very risky" (OR = 1.64, 95% CI = 1.08, 2.41) while occasional LCC smoking is only "somewhat risky" (OR = 1.71, 95% CI = 1.02, 2.87). Of the dual smokers, 20.7% intended to continue smoking LCCs in the future. Perceptions of addiction and risk of daily LCC smoking significantly predicted intention to continue LCC smoking. Addiction perceptions also significantly predicted susceptibility to initiate LCC smoking. Perceptions about harms from and addiction to LCCs could predict future LCC smoking. Health communication campaigns need to address the harms of LCCs. Our data suggest that perceptions of risk about the addictiveness of LCCs and frequency of use are important determinants of the LCC smoking susceptibility among some cigarette smokers and intended continued use among cigarette smokers with a history of LCC use. Health communication campaigns should address misperceptions related to LCCs.


Impaired postural stability in patients with cervical myelopathy: evaluation by computerized static stabilometry.
A prospective clinical study. To quantitatively evaluate impairment of postural stability in patients with cervical myelopathy. Proprioceptive sensation plays an important role in coordinated movement of the lower extremities and postural stability. Nevertheless, although disturbance of proprioceptive information will have an influence on the maintenance of postural stability, there have been few studies dealing with deterioration of postural stability in patients with cervical myelopathy. We investigated 52 cervical myelopathy patients who could stand without support and compared the results with those of 29 age-matched healthy volunteers. Postural stability was examined using a stabilometer. In the stabilometer, sway of gravity center was measured at upright position with eyes closed for 30 seconds. We used 2 parameters for evaluation: environmental area (EA), which measures degree of sway of the gravity center, and locus length per environmental area (L/EA), which measures fine control of standing posture by proprioceptive reflexes. The mean EA of the patient group was 13.9, whereas that of the control group was 2.74, revealing significantly larger postural instability in the patient group compared to the control group. The L/EA of the patient group was significantly worse than the control group. It was also shown that postural instability was significantly larger in the myelopathy patients with the severe clinical symptoms. The results of the present study demonstrate impairment of postural stability in patients with cervical myelopathy. The stabilometer can objectively evaluate the postural stability, which may reflect the function of the dorsal columns and the corticospinal tracts. Thus, stabilometry is a useful method for measuring a part of proprioceptive function and for objective assessment of the lower limb function of cervical myelopathy.


Generation of vortex waves in non-coaxial cylindrical waveguides.
A non-coaxial waveguide composed of a cylindrical resonator of radius R and cylindrical waveguides with the radii r1 and r2, respectively, is considered. The radii satisfy the inequality r1<r2<R. The conversion from the channel with zero orbital angular momentum (OAM) into the channels with non-zero OAM is achieved by shifting the center lines of the waveguides relative to the center line of the cylindrical resonator. The center lines of input and output waveguides are shifted relative to each other by the angle Δϕ in order to twist the output acoustic wave. The conversion efficiency of the input wave with zero OAM into the output wave with non-zero OAM as dependent on the frequency, length of the resonator, and Δϕ is considered, and the domains where the efficiency can reach almost 100% are found.


From Static to Dynamic: Electron Density of HOMO at Biaryl Linkage Controls the Mechanism of Hole Delocalization.
In order to extend the physical length of hole delocalization in a molecular wire, chromophores of increasing size are often desired. However, the effect of size on the efficacy and mechanism of hole delocalization remains elusive. Here, we employ a model set of biaryls to show that with increasing chromophore size, the mechanism of steady-state hole distribution switches from static delocalization in biaryls with smaller chromophores to dynamic hopping, as exemplified in the largest system, tBuHBC2 (i.e., "superbiphenyl"), which displays a vanishingly small electronic coupling. This important finding is analyzed with the aid of Hückel molecular orbital and Marcus-Hush theories. Our findings will enable the rational design of the novel molecular wires with length-invariant redox/optical properties suitable for long-range charge transfer.


From Body to Mind and Spirit: Qigong Exercise for Bereaved Persons with Chronic Fatigue Syndrome-Like Illness.
Bereavement may bring negative impacts on the mind, body, and spiritual well-being of grieving persons. Some bereaved persons with chronic fatigue syndrome- (CFS-) illness experience a dual burden of distress. This study investigated the effects of bereavement on CFS-like illness by comparing bereaved and nonbereaved participants. It also adopted a random group design to investigate the effectiveness of Qigong on improving the well-being of bereaved participants. The Qigong intervention comprised 10 group sessions delivered twice a week for 5 weeks and home-practice for at least three times a week lasting 15-30 minutes each. The participants' fatigue, anxiety, and depression, quality of life (QoL), and spiritual well-being were measured at baseline and 3 months after treatment. The bereaved participants experienced significantly greater mental fatigue (16.09 versus 14.44, p = 0.017) and lower physical QoL (34.02 versus 37.17, p = 0.011) than their nonbereaved counterparts. After 3 months, the mental fatigue (-8 versus -4, p = 0.010) and physical fatigue (-10 versus -5, p = 0.007) experienced by intervention group had declined significantly, and improvements on their spirituality (14 versus -2, p = 0.013) and psychological QoL (8.91 versus 0.69, p = 0.002) scores exceeded those of the control group.


Functional characteristics of cancer stem cells and their role in drug resistance of prostate cancer.
Cancer stem cells (CSCs) have the ability to self-renew and differentiate to give rise to heterogeneous phenotype of the tumor cells. It is believed that these cells are involved in metastasis, recurrence and therapy resistance in various cancers. CSCs have been identified in prostate cancer (PCa), one of the most diagnosed malignancies in men over the world, for which chemotherapy resistance is a major problem in the treatment of castration-resistant advanced stages. Molecular signatures, gene expression and functional features have been reported for PCa CSCs. Most data come from cell lines which may not represent the actual tumor. In the present work, a CSCs enriched population obtained from PCa explants was functionally characterized and analyzed for drug resistance. Tumorsphere cultures positive for ABCG2 transporter, CD133, CD44, cytokeratins 5 and 18 (CK5 and CK18) and negatives for androgen receptor (AR) and prostate-specific antigen (PSA) showed higher clonogenic capacity, holoclone-forming ability, colony-forming capacity in soft agar and lower proliferative and apoptotic rate than control adherent cell cultures. Furthermore, exposing tumorsphere cultures to ABCG2 substrate drugs resulted in a high survival rate compared with control PCa cells. This high drug resistance was decreased using a selective inhibitor of ABCG2. According to these results, tumorspheres from PCa explants showed a functional stem phenotype and a marked drug resistance, probably mediated by high expression of the ABCG2 transporter, which might be considered as a suitable therapeutic target for CSCs.


Desirable cytolytic immune effector cell recruitment by interleukin-15 dendritic cells.
Success of dendritic cell (DC) therapy in treating malignancies is depending on the DC capacity to attract immune effector cells, considering their reciprocal crosstalk is partially regulated by cell-contact-dependent mechanisms. Although critical for therapeutic efficacy, immune cell recruitment is a largely overlooked aspect regarding optimization of DC vaccination. In this paper we have made a head-to-head comparison of interleukin (IL)-15-cultured DCs and conventional IL-4-cultured DCs with regard to their proficiency in the recruitment of (innate) immune effector cells. Here, we demonstrate that IL-4 DCs are suboptimal in attracting effector lymphocytes, while IL15 DCs provide a favorable chemokine milieu for recruiting CD8+ T cells, natural killer (NK) cells and gamma delta (γδ) T cells. Gene expression analysis revealed that IL-15 DCs exhibit a high expression of chemokines involved in antitumor immune effector cell attraction, while IL-4 DCs display a more immunoregulatory profile characterized by the expression of Th2 and regulatory T cell-attracting chemokines. This is confirmed by functional data indicating an enhanced recruitment of granzyme B+ effector lymphocytes by IL-15 DCs, as compared to IL-4 DCs, and subsequent superior killing of tumor cells by the migrated lymphocytes. Elevated CCL4 gene expression in IL-15 DCs and lowered CCR5 expression on both migrated γδ T cells and NK cells, led to validation of increased CCL4 secretion by IL15 DCs. Moreover, neutralization of CCR5 prior to migration resulted in an important inhibition of γδ T cell and NK cell recruitment by IL-15 DCs. These findings further underscore the strong immunotherapeutic potential of IL-15 DCs.


Metabolomics in Sepsis and Its Impact on Public Health.
Sepsis, with its often devastating consequences for patients and their families, remains a major public health concern that poses an increasing financial burden. Early resuscitation together with the elucidation of the biological pathways and pathophysiological mechanisms with the use of "-omics" technologies have started changing the clinical and research landscape in sepsis. Metabolomics (i.e., the study of the metabolome), an "-omics" technology further down in the "-omics" cascade between the genome and the phenome, could be particularly fruitful in sepsis research with the potential to alter the clinical practice. Apart from its benefit for the individual patient, metabolomics has an impact on public health that extends beyond its applications in medicine. In this review, we present recent developments in metabolomics research in sepsis, with a focus on pneumonia, and we discuss the impact of metabolomics on public health, with a focus on free/libre open source software.


Ascorbate peroxidase. A prominent membrane protein in oilseed glyoxysomes.
The glyoxysomes of growing oilseed seedlings produce H2O2, a reactive oxygen species, during the beta-oxidation of lipids stored in the cotyledons. An expression library of dark-grown cotton (Gossypium hirsutm L.) cotyledons was screened with antibodies that recognized a 31-kD glyoxysomal membrane polypeptide. A full-length cDNA clone (1258 bp) was isolated that encodes a 32-kD subunit of ascorbate peroxidase (APX) with a single, putative membrane-spanning region near the C-terminal end of the polypeptide. Internal amino acid sequence analysis of the cotton 31-kD polypeptide verified that this clone encoded this protein. This enzyme, designated gmAPX, was immunocytochemically and enzymatically localized to the glyoxysomal membrane in cotton cotyledons. The activity of monodehydroascorbate reductase, a protein that reduces monodehydroascorbate to ascorbate with NADH, also was detected in these membranes. The co-localization of gmAPX and monodehydroascorbate reductase within the glyoxysomal membrane likely reflects an essential pathway for scavenging reactive oxygen species and also provides a mechanism to regenerate NAD+ for the continued operation of the glyoxylate cycle and beta-oxidation of fatty acids. Immunological cross-reactivity of 30- to 32-kD proteins in glyoxysomal membranes of cucumber, sunflower, castor bean, and cotton indicate that gmAPX is common among oilseed species.


Production of matrix metalloproteinases in response to mycobacterial infection.
Matrix metalloproteinases (MMPs) constitute a large family of enzymes with specificity for the various proteins of the extracellular matrix which are implicated in tissue remodeling processes and chronic inflammatory conditions. To investigate the role of MMPs in immunity to mycobacterial infections, we incubated murine peritoneal macrophages with viable Mycobacterium bovis BCG or Mycobacterium tuberculosis H37Rv and assayed MMP activity in the supernatants by zymography. Resting macrophages secreted only small amounts of MMP-9 (gelatinase B), but secretion increased dramatically in a dose-dependent manner in response to either BCG or M. tuberculosis in vitro. Incubation with mycobacteria also induced increased MMP-2 (gelatinase A) activity. Neutralization of tumor necrosis alpha (TNF-alpha), and to a lesser extent interleukin 18 (IL-18), substantially reduced MMP production in response to mycobacteria. Exogenous addition of TNF-alpha or IL-18 induced macrophages to express MMPs, even in the absence of bacteria. The immunoregulatory cytokines gamma interferon (IFN-gamma), IL-4, and IL-10 all suppressed BCG-induced MMP production, but through different mechanisms. IFN-gamma treatment increased macrophage secretion of TNF-alpha but still reduced their MMP activity. Conversely, IL-4 and IL-10 seemed to act by reducing the amount of TNF-alpha available to the macrophages. Finally, infection of BALB/c or severe combined immunodeficiency (SCID) mice with either BCG or M. tuberculosis induced substantial increases in MMP-9 activity in infected tissues. In conclusion, we show that mycobacterial infection induces MMP-9 activity both in vitro and in vivo and that this is regulated by TNF-alpha, IL-18, and IFN-gamma. These findings indicate a possible contribution of MMPs to tissue remodeling processes that occur in mycobacterial infections.


Effects of 2 Neonicotinoid Insecticides on Blood Cell Profiles and Corticosterone Concentrations of Wood Frogs (Lithobates sylvaticus).
Neonicotinoids are widely used insecticides that are detectable in agricultural waterways. These insecticides are of concern due to their potential impacts on nontarget organisms. Pesticides can affect development of amphibians and suppress the immune system, which could impact disease susceptibility and tolerance. No previous studies on amphibians have examined the effects of these insecticides on differential blood cell proportions or concentrations of corticosterone (a general stress hormone). We investigated the effects of chronic exposure to 2 neonicotinoids, thiamethoxam and clothianidin, on immunometrics of wood frogs (Lithobates sylvaticus). Frogs were exposed to single, chronic treatments of 2.5 or 250 µg/L of clothianidin or thiamethoxam for 7 wk from Gosner stages 25 to 46. The juvenile frogs were then maintained for 3 wk post metamorphosis without exposure to neonicotinoids. We measured water-borne corticosterone twice: at 6 d and 8 wk after exposure in larval and juvenile frogs, respectively. We assessed differential blood cell profiles from juvenile frogs. Corticosterone was significantly lower in tadpoles exposed to 250 µg/L of thiamethoxam compared with other tadpole treatments, but no significant differences in corticosterone concentrations were found in treatments using juvenile frogs. Anemia was detected in all treatments compared with controls with the exception of tadpoles exposed to 2.5 µg/L of clothianidin. Neutrophil-to-leukocyte and neutrophil-to-lymphocyte ratios were elevated in frogs exposed to 250 µg/L of thiamethoxam. Collectively, these results indicate that chronic exposure to neonicotinoids has varied impacts on blood cell profiles and corticosterone concentrations of developing wood frogs, which are indicative of stress. Future studies should investigate whether exposure to neonicotinoids increases susceptibility to infection by parasites in both larval and adult wood frogs. Environ Toxicol Chem 2019;38:1273-1284. © 2019 Crown in the right of Canada. Published by Wiley Periodicals Inc. on behalf of SETAC.


Effects of virion surface gp120 density on infection by HIV-1 and viral production by infected cells.
The quantity of envelope glycoprotein molecules (Env) on HIV-1 particles is still an issue of debate and, depending on the strain of virus and the nature of the producer cells, it can vary greatly. Here, we have attempted to address how Env density influences HIV-1 fitness. To this aim, we have produced HIV-1-derived viral particles with various amounts of R5 Env (low Env: Envlo; high Env: Envhi), using a regulatable expression system. The infectivity was assayed on human cells, engineered to express the HIV receptor CD4 and the co-receptor CCR5, as well as on peripheral blood lymphocytes and macrophages. In these experiments, low levels of Env were sufficient for cell infection, albeit at low efficiency. Increasing the amount of Env resulted in cooperatively improved infectivity, but a threshold was rapidly attained, indicating that only a fraction of Env was required for efficient infection. Unexpectedly, Env incorporation beyond what gives maximal infection transiently stimulated the expression of proviral genes, as well as retrovirus production, in newly infected cells. This was likely a consequence of induced NF-kappaB activity, as this transcription factor is triggered by Envhi, but not by Envlo, virions. Thus, our data suggest that one major effect of high Env density on the surface of HIV may not be better infection yields but rather improved viral production by newly infected cells.


Case Report: Testicular failure possibly associated with chronic use of methylphenidate.
Methylphenidate is a commonly prescribed treatment for attention deficit hyperactivity disorder (ADHD). However, little is known about its adverse effects on the male reproductive system. We report a 20-year-old male patient whose chief complaint was of delayed puberty. He spoke in a high-pitched voice and complained of lack of body hair, impaired libido, inadequate erectile function, chronic fatigue, and low energy. He had been treated with methylphenidate as an infant and had continued treatment for 17 years. On examination, the patient was lean and visibly lacked facial or body hair. He further explained that he had never been able to grow underarm or facial hair and that he was often mistakenly considered a young teenager rather than a 20-year-old. The patient's genitalia were categorized as Tanner Stage 2. Laboratory studies confirmed low serum follicle-stimulating hormone (FSH), luteinizing hormone (LH), and testosterone levels. The patient was given exogenous testosterone supplementation with pellets and human chorionic gonadotropin to maintain testicular size. After 4 months his symptoms improved and he demonstrated signs of puberty. Our goal is to further elucidate the possible impact of methylphenidate on the male reproductive system.


Synthesis, structure and reactivity of Pd and Ir complexes based on new lutidine-derived NHC/phosphine mixed pincer ligands.
Coordination studies of new lutidine-derived hybrid NHC/phosphine ligands (CNP) to Pd and Ir have been performed. Treatment of the square-planar [Pd(CNP)Cl](AgCl2) complex 2a with KHMDS produces the selective deprotonation at the CH2P arm of the pincer to yield the pyridine-dearomatised complex 3a. A series of cationic [Ir(CNP)(cod)]+ complexes 4 has been prepared by reaction of the imidazolium salts 1 with Ir(acac)(cod). These derivatives exhibit in the solid state, and in solution, a distorted trigonal bipyramidal structure in which the CNP ligands adopt an unusual C(axial)-N(equatorial)-P(equatorial) coordination mode. Reactions of complexes 4 with CO and H2 yield the carbonyl species 5a(Cl) and 6a(Cl), and the dihydrido derivatives 7, respectively. Furthermore, upon reaction of complex 4b(Br) with base, selective deprotonation at the methylene CH2P arms is observed. The, thus formed, deprotonated Ir complex 8b reacts with H2 in a ligand-assisted process leading to the trihydrido complex 9b, which can also be obtained by reaction of 7b(Cl) with H2 in the presence of KOtBu. Finally, the catalytic activity of Ir-CNP complexes in the hydrogenation of ketones has been briefly assessed.


Characterization of Epigallocatechin-Gallate-Grafted Chitosan Nanoparticles and Evaluation of Their Antibacterial and Antioxidant Potential.
Nanoparticles based on chitosan modified with epigallocatechin gallate (EGCG) were synthetized by nanoprecipitation (EGCG-g-chitosan-P). Chitosan was modified by free-radical-induced grafting, which was verified by Fourier transform infrared (FTIR). Furthermore, the morphology, particle size, polydispersity index, and zeta potential of the nanoparticles were investigated. The grafting degree of EGCG, reactive oxygen species (ROS) production, antibacterial and antioxidant activities of EGCG-g-chitosan-P were evaluated and compared with those of pure EGCG and chitosan nanoparticles (Chitosan-P). FTIR results confirmed the modification of the chitosan with EGCG. The EGCG-g-chitosan-P showed spherical shapes and smoother surfaces than those of Chitosan-P. EGCG content of the grafted chitosan nanoparticles was 330 μg/g. Minimal inhibitory concentration (MIC) of EGCG-g-chitosan-P (15.6 μg/mL) was lower than Chitosan-P (31.2 μg/mL) and EGCG (500 μg/mL) against Pseudomonas fluorescens (p < 0.05). Additionally, EGCG-g-chitosan-P and Chitosan-P presented higher Staphylococcus aureus growth inhibition (100%) than EGCG at the lowest concentration tested. The nanoparticles produced an increase of ROS (p < 0.05) in both bacterial species assayed. Furthermore, EGCG-g-chitosan-P exhibited higher antioxidant activity than that of Chitosan-P (p < 0.05) in 2,2'-azino-bis (3-ethyl-benzothiazoline-6-sulfonic acid) (ABTS), 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) and ferric-reducing antioxidant power assays. Based on the above results, EGCG-g-chitosan-P shows the potential for food packaging and biomedical applications.


A noncanonical E-box enhancer drives mouse Period2 circadian oscillations in vivo.
The mouse Period2 (mPer2) locus is an essential negative-feedback element of the mammalian circadian-clock mechanism. Recent work has shown that mPer2 circadian gene expression persists in both central and peripheral tissues. Here, we analyze the mouse mPer2 promoter and identify a circadian enhancer (E2) with a noncanonical 5'-CACGTT-3' E-box located 20 bp upstream of the mPer2 transcription start site. The E2 enhancer accounts for most circadian transcriptional drive of the mPer2 locus by CLOCK:BMAL1, is a major site of DNaseI hypersensitivity in this region, and is constitutively bound by a transcriptional complex containing the CLOCK protein. Importantly, the E2 enhancer is sufficient to drive self-sustained circadian rhythms of luciferase activity in central and peripheral tissues from mPer2-E2::Luciferase transgenic mice with tissue-specific phase and period characteristics. Last, genetic analysis with mutations in Clock and Bmal1 shows that the E2 enhancer is a target of CLOCK and BMAL1 in vivo.


Multivalent Flexible Nanogels Exhibit Broad-Spectrum Antiviral Activity by Blocking Virus Entry.
The entry process of viruses into host cells is complex and involves stable but transient multivalent interactions with different cell surface receptors. The initial contact of several viruses begins with attachment to heparan sulfate (HS) proteoglycans on the cell surface, which results in a cascade of events that end up with virus entry. The development of antiviral agents based on multivalent interactions to shield virus particles and block initial interactions with cellular receptors has attracted attention in antiviral research. Here, we designed nanogels with different degrees of flexibility based on dendritic polyglycerol sulfate to mimic cellular HS. The designed nanogels are nontoxic and broad-spectrum, can multivalently interact with viral glycoproteins, shield virus surfaces, and efficiently block infection. We also visualized virus-nanogel interactions as well as the uptake of nanogels by the cells through clathrin-mediated endocytosis using confocal microscopy. As many human viruses attach to the cells through HS moieties, we introduce our flexible nanogels as robust inhibitors for these viruses.


Mu element-generated gene conversions in maize attenuate the dominant knotted phenotype.
The knotted1 gene was first defined by dominant mutations that affect leaf morphology. The original allele, Kn1-O, results from a 17-kb tandem duplication. Mutator (Mu) insertions near the junction of the two repeats suppress the leaf phenotype to different degrees depending on the position of the insertion. The Mu insertions also increase the frequency of recombination at Kn1-O to create derivative alleles in which the Mu element and one copy of the repeat are lost. These derivatives are normal in appearance. Here we describe two derivatives that retained the tandem duplication but gained insertions of 1.7 and 3 kb in length in place of the Mu element. In each case, the inserted DNA is a sequence that normally flanks the distal repeat unit. Thus, each derivative consists of a tandem duplication in which the repeat unit has been extended at its distal end by the length of the new insertion. The 1.7-kb insertion dampens the phenotype, as did the original Mu insertion, whereas the 3-kb insertion completely suppresses the knotted phenotype. We propose that gene conversion, stimulated by the double-strand break of the Mu excision, gave rise to these derivatives.


Cellular Senescence: Defining a Path Forward.
Cellular senescence is a cell state implicated in various physiological processes and a wide spectrum of age-related diseases. Recently, interest in therapeutically targeting senescence to improve healthy aging and age-related disease, otherwise known as senotherapy, has been growing rapidly. Thus, the accurate detection of senescent cells, especially in vivo, is essential. Here, we present a consensus from the International Cell Senescence Association (ICSA), defining and discussing key cellular and molecular features of senescence and offering recommendations on how to use them as biomarkers. We also present a resource tool to facilitate the identification of genes linked with senescence, SeneQuest (available at http://Senequest.net). Lastly, we propose an algorithm to accurately assess and quantify senescence, both in cultured cells and in vivo.


Optimum frequency of exercise for bone health: randomised controlled trial of a high-impact unilateral intervention.
Exercise can increase bone strength, but to be effective in reducing fracture risk, exercise must be feasible enough to be adopted into daily life and influence potentially vulnerable skeletal sites such as the superolateral cortex of the femoral neck, where thinning is associated with increased fracture risk. Brief, high-impact exercise increases femoral neck bone density but the optimal frequency of such exercise and the location of bone accrual is unknown. This study thus examined (1) the effectiveness of different weekly frequencies of exercise on femoral neck BMD and (2) whether BMD change differed between hip sites using a high-impact, unilateral intervention. Healthy premenopausal women were randomly assigned to exercise 0, 2, 4, or 7 days/week for 6 months. The exercise intervention incorporated 50 multidirectional hops on one randomly selected leg. BMD was measured by DXA at baseline and after 6 months of exercise. Changes in the exercise leg were compared between groups using ANCOVA, with change in the control leg and baseline BMD as covariates. RM-MANOVA was conducted to determine whether bone changes from exercise differed between hip sites. 61 women (age 33.6+/-11.1 years) completed the intervention. Compliance amongst exercisers was 86.7+/-10.6%. Peak ground reaction forces during exercise increased from 2.5 to 2.8 times body weight. The change in femoral neck BMD in the exercise limb (adjusted for change in the control limb and baseline BMD) differed between groups (p=0.015), being -0.3% (-1.2 to 0.6), 0.0% (-1.0 to 1.0), 0.9% (-0.1 to 2.0) and 1.8% (0.8 to 2.8) in those exercising 0, 2, 4 and 7 days per week, respectively. When BMD changes at upper neck, lower neck and trochanter were compared using RM-MANOVA, a significant exercise effect was observed (p=0.048), but this did not differ significantly between sites (p=0.439) despite greatest mean increases at the upper femoral neck. Brief, daily hopping exercises increased femoral neck BMD in premenopausal women but less frequent exercise was not effective. Brief high-impact exercise may have a role in reducing hip fragility, but may need to be performed frequently for optimal response.


Predicting emotional and social competence during early childhood from toddler risk and maternal behavior.
The longitudinal associations between maternal parenting behavior and toddler risk with children's emotional and social competence were examined during the transition to kindergarten, in a sample of 253 children. Toddler risk was characterized by early externalizing behavior and poor emotion regulation skills. Given that we were interested in the multiple pathways that may result in emotional and social competence, we examined the interactions among maternal parenting behavior and toddler risk. There were some significant interactions, although the pattern of results was not consistent across all competence outcomes. Maternal parenting behavior was not directly associated with children's emotional and social competence. In some instances, maternal control has differential implications for children's emotional and social competence dependent upon the child's level of early risk and maternal positive parenting. Specifically, maternal control tended to be more detrimental for children's emotional competence during the transition to kindergarten, when children exhibit higher levels of risk. Overall, it appears that there are multiple developmental pathways, depending on child and maternal characteristics that lead to early emotional and social competence.


Regulatory T cells may participate in Helicobacter pylori persistence in gastric MALT lymphoma: lessons from an animal model.
It has been postulated that the emergence of autoimmune gastritis in neonatal thymectomised (d3Tx) BALB/c mice may be a consequence of post-surgery deficit in Tregs. In this study, previously obtained samples from d3Tx mice were used in order to determine whether thymectomy creates a deficit in this T cell subset thereby allowing the emergence of autoimmune phenomena as a prerequisite for GML. The splenic Treg reserve and the local recruitment of these cells in the gastric mucosa were investigated using complementary molecular and immunohistochemistry approaches. Higher Foxp3/CD3 ratios were found in the spleen of non-infected d3Tx mice compared to non-thymectomised (NTx) controls. These results indicate a relative enrichment of Tregs following thymectomy in adult mice. The absence of Treg depletion in d3Tx mice is in line with the absence of auto-immune gastritis in non-infected d3Tx mice. Higher levels of T cell and Treg infiltration were also found in the stomach of GML-developing d3Tx mice versus NTx mice. Surprisingly, inflammatory scores inversely correlated with the bacterial inoculum. The presence of a small Treg containing compartment among gastric biopsies of GML developing d3Tx mice may play a role in perseverance of a minimal bacterial numbers thereby maintaining an antigen-dependent stimulation and proliferation.


The Use of Social Media by Dental Students for Communication and Learning: Two Viewpoints: Viewpoint 1: Social Media Use Can Benefit Dental Students' Communication and Learning and Viewpoint 2: Potential Problems with Social Media Outweigh Their Benefits for Dental Education.
Social media have become a major part of an interconnected society, impacting personal and professional lives. This Point/Counterpoint presents two opposing viewpoints on the question of whether social media should be used in dental education as a learning and communication tool for dental students. Viewpoint 1 argues that social media benefit student learning and should be used as a tool in dental education. This argument is based on evidence concerning use of social media and improved learning across health professions, improved peer-peer communication in clinical education, improved engagement in interprofessional education (IPE), and provision of a mechanism for safe and improved communication between practitioners and patients, as well as faculty and students. Viewpoint 2 argues that potential problems and risks in using social media outweigh any benefits found in learning and therefore social media should not be used as a tool in dental education. This viewpoint is supported by evidence of negative effects on learning, the establishment of a negative digital footprint in the public's view, risk of privacy violations when using social media, and the new phenomenon of Internet addiction with its negative physiological effects on social media users.


Dynamics of infection in tick vectors and at the tick-host interface.
Tick-borne flaviviruses are common, widespread, and successfully adapted to their mode of transmission. Most tick vectors of flaviviruses are ixodid species. These ticks are characterized by a comparatively long life cycle, lasting several years, during which the infecting virus may be maintained from one developmental stage of the tick to the next. Hence ticks act as highly efficient reservoirs of flaviviruses. Many tick-borne flaviviruses are transmitted vertically, from adult to offspring, although the frequency is too low to maintain the viruses solely in the tick population. Instead, the survival of tick-borne flaviviruses is dependent on horizontal transmission, both from an infected tick to a susceptible vertebrate host and from an infected vertebrate to uninfected ticks feeding on the animal. The dynamics of transmission and infection have traditionally been considered in isolation: in the tick, following virus uptake in the infected blood meal, infection of the midgut, passage through the hemocoel to the salivary glands, and transmission via the saliva; and in the vertebrate host, virus delivery into the skin at the site of tick feeding, infection of the draining lymph nodes, and dissemination to target organs. However, there is now compelling evidence of a complex interaction between the tick vector and its vertebrate host that affects virus transmission profoundly. The feeding site in the skin is a battleground in which the hemostatic, inflammatory, and immune responses of the host are countered by antihemostatic, anti-inflammatory, and immunomodulatory molecules (mostly proteins and peptides) secreted in tick saliva. Here we speculate that exploitation of the tick pharmacopeia, rather than development of viremia, is the key step in successful tick-borne flavivirus transmission.


Structural basis of HIV-1 neutralization by affinity matured Fabs directed against the internal trimeric coiled-coil of gp41.
The conserved internal trimeric coiled-coil of the N-heptad repeat (N-HR) of HIV-1 gp41 is transiently exposed during the fusion process by forming a pre-hairpin intermediate, thus representing an attractive target for the design of fusion inhibitors and neutralizing antibodies. In previous studies we reported a series of broadly neutralizing mini-antibodies derived from a synthetic naïve human combinatorial antibody library by panning against a mimetic of the trimeric N-HR coiled coil, followed by affinity maturation using targeted diversification of the CDR-H2 loop. Here we report crystal structures of the N-HR mimetic 5-Helix with two Fabs that represent the extremes of this series: Fab 8066 is broadly neutralizing across a wide panel of B and C type HIV-1 viruses, whereas Fab 8062 is non-neutralizing. The crystal structures reveal important differences in the conformations of the CDR-H2 loops in the complexes that propagate into other regions of the antigen-antibody interface, and suggest that both neutralization properties and affinity for the target can be attributed, at least in part, to the differences in the interactions of the CDR-H2 loops with the antigen. Furthermore, modeling of the complex of an N-HR trimer with three Fabs suggests that the CDR-H2 loop may be involved in close intermolecular contacts between neighboring antibody molecules, and that such contacts may hinder the formation of complexes between the N-HR trimer and more than one antibody molecule depending on the conformation of the bound CDR-H2 loop which is defined by its interactions with antigen. Comparison with the crystal structure of the complex of 5-Helix with another neutralizing monoclonal antibody known as D5, derived using an entirely different antibody library and panning procedure, reveals remarkable convergence in the optimal sequence and conformation of the CDR-H2 loop.


CDK prevents Mcm2-7 helicase loading by inhibiting Cdt1 interaction with Orc6.
In Saccharomyces cerevisiae cells, B-type cyclin-dependent kinases (CDKs) target two origin recognition complex (ORC) subunits, Orc2 and Orc6, to inhibit helicase loading. We show that helicase loading by ORC is inhibited by two distinct CDK-dependent mechanisms. Independent of phosphorylation, binding of CDK to an "RXL" cyclin-binding motif in Orc6 sterically reduces the initial recruitment of the Cdt1/Mcm2-7 complex to ORC. CDK phosphorylation of Orc2 and Orc6 inhibits the same step in helicase loading. This phosphorylation of Orc6 is stimulated by the RXL motif and mediates the bulk of the phosphorylation-dependent inhibition of helicase loading. Direct binding experiments show that CDK phosphorylation specifically blocks one of the two Cdt1-binding sites on Orc6. Consistent with the inactivation of one Cdt1-binding site preventing helicase loading, CDK phosphorylation of ORC causes a twofold reduction of initial Cdt1/Mcm2-7 recruitment but results in nearly complete inhibition of Mcm2-7 loading. Intriguingly, in addition to being a target of both CDK inhibitory mechanisms, the Orc6 RXL/cyclin-binding motif plays a positive role in the initial recruitment of Cdt1/Mcm2-7 to the origin, suggesting that this motif is critical for the switch between active and inhibited ORC function at the G1-to-S-phase transition.


Comprehensive microRNA profiling reveals a unique human embryonic stem cell signature dominated by a single seed sequence.
Embryonic stem cells are unique among cultured cells in their ability to self-renew and differentiate into a wide diversity of cell types, suggesting that a specific molecular control network underlies these features. Human embryonic stem cells (hESCs) are known to have distinct mRNA expression, global DNA methylation, and chromatin profiles, but the involvement of high-level regulators, such as microRNAs (miRNA), in the hESC-specific molecular network is poorly understood. We report that global miRNA expression profiling of hESCs and a variety of stem cell and differentiated cell types using a novel microarray platform revealed a unique set of miRNAs differentially regulated in hESCs, including numerous miRNAs not previously linked to hESCs. These hESC-associated miRNAs were more likely to be located in large genomic clusters, and less likely to be located in introns of coding genes. hESCs had higher expression of oncogenic miRNAs and lower expression of tumor suppressor miRNAs than the other cell types. Many miRNAs upregulated in hESCs share a common consensus seed sequence, suggesting that there is cooperative regulation of a critical set of target miRNAs. We propose that miRNAs are coordinately controlled in hESCs, and are key regulators of pluripotence and differentiation. Disclosure of potential conflicts of interest is found at the end of this article.


Exploring the capacity of minimalist protein interfaces: interface energetics and affinity maturation to picomolar KD of a single-domain antibody with a flat paratope.
A major architectural class in engineered binding proteins ("antibody mimics") involves the presentation of recognition loops off a single-domain scaffold. This class of binding proteins, both natural and synthetic, has a strong tendency to bind a preformed cleft using a convex binding interface (paratope). To explore their capacity to produce high-affinity interfaces with diverse shape and topography, we examined the interface energetics and explored the affinity limit achievable with a flat paratope. We chose a minimalist paratope limited to two loops found in a natural camelid heavy-chain antibody (VHH) that binds to ribonuclease A. Ala scanning of the VHH revealed only three "hot spot" side chains and additional four residues important for supporting backbone-mediated interactions. The small number of critical residues suggested that this is not an optimized paratope. Using selection from synthetic combinatorial libraries, we enhanced its affinity by >100-fold, resulting in variants with Kd as low as 180 pM with no detectable loss of binding specificity. High-resolution crystal structures revealed that the mutations induced only subtle structural changes but extended the network of interactions. This resulted in an expanded hot spot region including four additional residues located at the periphery of the paratope with a concomitant loss of the so-called "O-ring" arrangement of energetically inert residues. These results suggest that this class of simple, single-domain scaffolds is capable of generating high-performance binding interfaces with diverse shape. More generally, they suggest that highly functional interfaces can be designed without closely mimicking natural interfaces.


Predicting HIV-1 transmission and antibody neutralization efficacy in vivo from stoichiometric parameters.
The potential of broadly neutralizing antibodies targeting the HIV-1 envelope trimer to prevent HIV-1 transmission has opened new avenues for therapies and vaccines. However, their implementation remains challenging and would profit from a deepened mechanistic understanding of HIV-antibody interactions and the mucosal transmission process. In this study we experimentally determined stoichiometric parameters of the HIV-1 trimer-antibody interaction, confirming that binding of one antibody is sufficient for trimer neutralization. This defines numerical requirements for HIV-1 virion neutralization and thereby enables mathematical modelling of in vitro and in vivo antibody neutralization efficacy. The model we developed accurately predicts antibody efficacy in animal passive immunization studies and provides estimates for protective mucosal antibody concentrations. Furthermore, we derive estimates of the probability for a single virion to start host infection and the risks of male-to-female HIV-1 transmission per sexual intercourse. Our work thereby delivers comprehensive quantitative insights into both the molecular principles governing HIV-antibody interactions and the initial steps of mucosal HIV-1 transmission. These insights, alongside the underlying, adaptable modelling framework presented here, will be valuable for supporting in silico pre-trial planning and post-hoc evaluation of HIV-1 vaccination or antibody treatment trials.


Prognostic significance of erythropoietin in pancreatic adenocarcinoma.
Erythropoietin (Epo) administration has been reported to have tumor-promoting effects in anemic cancer patients. We investigated the prognostic impact of endogenous Epo in patients with pancreatic ductal adenocarcinoma (PDAC). The clinico-pathological relevance of hemoglobin (Hb, n = 150), serum Epo (sEpo, n = 87) and tissue expression of Epo/Epo receptor (EpoR, n = 104) was analyzed in patients with PDAC. Epo/EpoR expression, signaling, growth, invasion and chemoresistance were studied in Epo-exposed PDAC cell lines. Compared to donors, median preoperative Hb levels were reduced by 15% in both chronic pancreatitis (CP, p<0.05) and PDAC (p<0.001), reaching anemic grade in one third of patients. While inversely correlating to Hb (r = -0.46), 95% of sEPO values lay within the normal range. The individual levels of compensation were adequate in CP (observed to predicted ratio, O/P = 0.99) but not in PDAC (O/P = 0.85). Strikingly, lower sEPO values yielding inadequate Epo responses were prominent in non-metastatic M0-patients, whereas these parameters were restored in metastatic M1-group (8 vs. 13 mU/mL; O/P = 0.82 vs. 0.96; p<0.01)--although Hb levels and the prevalence of anemia were comparable. Higher sEpo values (upper quartile ≥ 16 mU/ml) were not significantly different in M0 (20%) and M1 (30%) groups, but were an independent prognostic factor for shorter survival (HR 2.20, 10 vs. 17 months, p<0.05). The pattern of Epo expression in pancreas and liver suggested ectopic release of Epo by capillaries/vasa vasorum and hepatocytes, regulated by but not emanating from tumor cells. Epo could initiate PI3K/Akt signaling via EpoR in PDAC cells but failed to alter their functions, probably due to co-expression of the soluble EpoR isoform, known to antagonize Epo. Higher sEPO levels counteract anemia but worsen outcome in PDAC patients. Further trials are required to clarify how overcoming a sEPO threshold ≥16 mU/ml by endogenous or exogenous means may predispose to or promote metastatic progression.


The effect of subclinical mastitis on milk yield in dairy goats.
The aims of this study were to estimate milk yield (MY) losses associated with subclinical intramammary infection (IMI) in dairy goats and to assess if somatic cell count (SCC) can be used to estimate such MY losses. We used 2 data sets to study these questions. The first data set consisted of 5 herds. Milk production and SCC were recorded during 1 lactation. From approximately 100 does in each herd, milk samples were collected on 3 occasions during lactation for bacteriological culture. Linear mixed regression was used to estimate the effect of IMI on MY. The second data set consisted of 6 large herds, in which some of the goats had an extended lactation (≥2 yr). Milk yield and SCC data were recorded without bacteriological culture. The data showed that bacterial infection was related to an increase in SCC. Infections with major pathogens were rare and associated with a decreased MY; infection with coagulase-negative staphylococci did not affect MY, whereas infection with Corynebacterium bovis was associated with increased MY. A negative correlation was observed between SCC and MY, but the data suggested that this negative correlation was attenuated rather than caused by IMI. Furthermore, SCC seemed to be affected by MY via a dilution effect. Hypotheses about biological mechanisms behind these observations are discussed. This paper shows that MY losses caused by subclinical udder infections are limited in goats, and that SCC cannot be used to estimate the magnitude of these losses.


Disease model curation improvements at Mouse Genome Informatics.
Optimal curation of human diseases requires an ontology or structured vocabulary that contains terms familiar to end users, is robust enough to support multiple levels of annotation granularity, is limited to disease terms and is stable enough to avoid extensive reannotation following updates. At Mouse Genome Informatics (MGI), we currently use disease terms from Online Mendelian Inheritance in Man (OMIM) to curate mouse models of human disease. While OMIM provides highly detailed disease records that are familiar to many in the medical community, it lacks structure to support multilevel annotation. To improve disease annotation at MGI, we evaluated the merged Medical Subject Headings (MeSH) and OMIM disease vocabulary created by the Comparative Toxicogenomics Database (CTD) project. Overlaying MeSH onto OMIM provides hierarchical access to broad disease terms, a feature missing from the OMIM. We created an extended version of the vocabulary to meet the genetic disease-specific curation needs at MGI. Here we describe our evaluation of the CTD application, the extensions made by MGI and discuss the strengths and weaknesses of this approach. DATABASE URL: http://www.informatics.jax.org/


Stakeholder's perspective: Sustainability of a community health worker program in Afghanistan.
The objectives of this study were two-fold: 1) to examine how different stakeholders define sustainability, and 2) to identify barriers to and facilitators of the sustainability of the Afghan CHW program. We interviewed 63 individual key informants, and conducted 11 focus groups [35 people] with policymakers, health managers, community health workers, and community members across Afghanistan. The participants were purposefully selected to provide a wide range of perspectives. Different stakeholders define sustainability differently. Policymakers emphasize financial resources; health managers, organizational operations; and community-level stakeholders, routine frontline activities. The facilitators they identify include integration into the health system, community support, and capable human resources. Barriers they noted include lack of financial resources, poor program design and implementation, and poor quality of services. Measures to ensure sustainability could be national revenue allocation, health-specific taxation, and community financing. Sustainability is complicated and has multiple facets. The plurality of understanding of sustainability among stakeholders should be addressed explicitly in the program design. To ensure sustainability, there is a need for a coordinated effort amongst all stakeholders.


Streptococcus gallolyticus infection in colorectal cancer and association with biological and clinical factors.
There is an unambiguous association of Streptococcus gallolyticus infection with colorectal cancer, although there is limited information about epidemiology or interaction between molecular and environmental factors. We performed an original quantitative analysis of S. gallolyticus in unselected colorectal cancer patients (n = 190) and their association with clinical, pathological tumor molecular profiles (microsatellite instability, hypermethylator phenotype and chromosomal instability pathways), and other biological factors in colorectal tumor and normal tissues (cytomegalovirus and Epstein-Barr virus infection). We developed a new quantitative method to assess bacterial load. Analytical validation was reached with a very high sensitivity and specificity. Our results showed a 3.2% prevalence of S. gallolyticus infection in our unselected cohort of colorectal cancer cases (6/190). The average S. gallolyticus copy number was 7,018 (range 44-34,585). No previous reports relating to S. gallolyticus infection have been published for unselected cohorts of patients. Finally, and despite a low prevalence of S. gallolyticus in this study, we were able to define a specific association with tumor tissue (p = 0.03) and with coinfection with Epstein-Barr virus (p = 0.042; OR: 9.49; 95% IC: 1.1-82.9). The prevalence data provided will be very useful in the design of future studies, and will make it possible to estimate the sample size needed to assess precise objectives. In conclusion, our results show a low prevalence of S. gallolyticus infection in unselected colorectal cancer patients and an association of positive S. gallolyticus infection with tumor tissue and Epstein-Barr virus coinfection. Further studies will be needed to definitively assess the prevalence of S. gallolyticus in colorectal cancer and the associated clinicopathological and molecular profiles.


Histaminergic descending inputs to the mesopontine tegmentum and their role in the control of cortical activation and wakefulness in the cat.
We have demonstrated previously the importance of histaminergic neurons in arousal mechanisms. In addition to their ascending axons, these neurons also send heavy descending inputs to the mesopontine tegmentum (MPT), which plays a key role in cortical activation during wakefulness (W). This anatomical link suggests histaminergic control of the mechanisms of the MPT relevant to behavioral states. In this study, we sought to demonstrate, at the light microscopy level, hypothalamotegmental histaminergic pathways and their topographical interaction with MPT neurons in the cat and to explore further their involvement in sleep-wake control. Using immunohistochemistry of histamine (HA), either alone or together with that of choline-acetyltransferase or tyrosine hydroxylase, a large number of very fine, short and varicose HA-positive fibers and terminal-like dots were detected in the MPT, including the laterodorsal tegmental nucleus, locus coeruleus (LC), LC alpha, and peri-LC alpha. Furthermore, these fibers and terminal-like structures were found in close proximity to a great number of cholinergic or noradrenergic neurons. We also investigated the effects of microadministration of HA agonists and antagonist into the mediodorsal pontine tegmentum on the cortical electroencephalogram (EEG) power spectra and the sleep-wake cycle in freely moving cats. Microinjection of HA or 2-thiazolylethylamine (an H1-receptor agonist) caused a long-lasting suppression of cortical slow activity and an increase in quiet wakefulness (W). Paradoxical sleep, however, was less affected. The effects of HA were attenuated by systemic or in situ pretreatment with mepyramine (an H1-receptor antagonist), which when injected alone produced an increase in slow wave sleep. Microinjection of impromidine (an H2-receptor agonist) into the same area had no effect on either the cortical EEG or W. Because MPT ascending and presumed cholinergic neurons discharge tonically during cortical activation of W and because HA causes excitation of MPT cholinergic neurons via H1 receptors, we hypothesize that the histaminergic descending afferents in the MPT would promote cortical desynchronization and W, at least partially, via activation of H1 receptors situated on cholinergic neurons and that the interactions between histaminergic and cholinergic neurons constitute an important circuit in cortical activation during W.


A deficit survival analysis to assess the natural history of uveal melanoma.
A deficit survival analysis was conducted using a retrospective series of 230 patients with uveal melanoma. All underwent enucleation as primary treatment for their disease. Considering the three leading prognostic factors (epithelioid cells per high power field, tumor size, and tumor location), median age was plotted against median survival to determine pathways of disease spread, local and metastatic disease phases, consequences of delayed diagnosis, and the timing of enucleation. The subgroup with all three factors favorable could be subdivided into younger and older subsets on the basis of a bimodal age distribution. Only the older subset could be identified as a local disease state where survival following enucleation corresponded to normal population survival. All remaining subgroups were metastatic because of survival deficits compared to the normal population. Disease tended to progress according to three well defined pathways with 6-8 years required to progress from one subgroup to the next along these paths. These pathways were characterized by constant deficit survivals, suggesting that delays in diagnosis did not translate into loss in survival. Moreover, the value of enucleation as primary therapy can be questioned in the context that it does not appear to alter the natural history of the disease, except for tumors greater than 10 mm in largest dimension which are located anterior to the equator with fewer than 2 epithelioid cells per high power field. Findings suggest that uveal melanoma can be treated by means other than enucleation to allow a chance for prolonged survival with vision preservation.


New approaches to the study of human brain networks underlying spatial attention and related processes.
Cognitive processes, such as spatial attention, are thought to rely on extended networks in the human brain. Both clinical data from lesioned patients and fMRI data acquired when healthy subjects perform particular cognitive tasks typically implicate a wide expanse of potentially contributing areas, rather than just a single brain area. Conversely, evidence from more targeted interventions, such as transcranial magnetic stimulation (TMS) or invasive microstimulation of the brain, or selective study of patients with highly focal brain damage, can sometimes indicate that a single brain area may make a key contribution to a particular cognitive process. But this in turn raises questions about how such a brain area may interface with other interconnected areas within a more extended network to support cognitive processes. Here, we provide a brief overview of new approaches that seek to characterise the causal role of particular brain areas within networks of several interacting areas, by measuring the effects of manipulations for a targeted area on function in remote interconnected areas. In human participants, these approaches include concurrent TMS-fMRI and TMS-EEG, as well as combination of the focal lesion method in selected patients with fMRI and/or EEG measures of the functional impact from the lesion on interconnected intact brain areas. Such approaches shed new light on how frontal cortex and parietal cortex modulate sensory areas in the service of attention and cognition, for the normal and damaged human brain.


Long-Term Behavioral Outcomes after a Randomized, Clinical Trial of Counselor-Assisted Problem Solving for Adolescents with Complicated Mild-to-Severe Traumatic Brain Injury.
Family problem-solving therapy (FPST) has been shown to reduce behavior problems after pediatric traumatic brain injury (TBI). It is unclear whether treatment gains are maintained. We sought to evaluate the maintenance of improvements in behavior problems after a Web-based counselor-assisted FPST (CAPS) intervention compared to an Internet resource comparison (IRC) intervention provided to adolescents within the initial year post-TBI. We hypothesized that family socioeconomic status, child educational status, and baseline levels of symptoms would moderate the efficacy of the treatment over time. Participants included 132 adolescents ages 12-17 years who sustained a complicated mild-to-severe TBI 1-6 months before study enrollment. Primary outcomes were the Child Behavior Checklist Internalizing and Externalizing Totals. Mixed-models analyses, using random intercepts and slopes, were conducted to examine group differences over time. There was a significant group×time×grade interaction (F(1,304)=4.42; p=0.03) for internalizing problems, with high school-age participants in CAPS reporting significantly lower symptoms at 18 months postbaseline than those in the IRC. Post-hoc analyses to elucidate the nature of effects on internalizing problems revealed significant group×time×grade interactions for the anxious/depressed (p=0.03) and somatic complaints subscales (p=0.04). Results also indicated significant improvement over time for CAPS participants who reported elevated externalizing behavior problems at baseline (F(1, 310)=7.17; p=0.008). Findings suggest that CAPS may lead to long-term improvements in behavior problems among older adolescents and those with pretreatment symptoms.


APP binds DR6 to trigger axon pruning and neuron death via distinct caspases.
Naturally occurring axonal pruning and neuronal cell death help to sculpt neuronal connections during development, but their mechanistic basis remains poorly understood. Here we report that beta-amyloid precursor protein (APP) and death receptor 6 (DR6, also known as TNFRSF21) activate a widespread caspase-dependent self-destruction program. DR6 is broadly expressed by developing neurons, and is required for normal cell body death and axonal pruning both in vivo and after trophic-factor deprivation in vitro. Unlike neuronal cell body apoptosis, which requires caspase 3, we show that axonal degeneration requires caspase 6, which is activated in a punctate pattern that parallels the pattern of axonal fragmentation. DR6 is activated locally by an inactive surface ligand(s) that is released in an active form after trophic-factor deprivation, and we identify APP as a DR6 ligand. Trophic-factor deprivation triggers the shedding of surface APP in a beta-secretase (BACE)-dependent manner. Loss- and gain-of-function studies support a model in which a cleaved amino-terminal fragment of APP (N-APP) binds DR6 and triggers degeneration. Genetic support is provided by a common neuromuscular junction phenotype in mutant mice. Our results indicate that APP and DR6 are components of a neuronal self-destruction pathway, and suggest that an extracellular fragment of APP, acting via DR6 and caspase 6, contributes to Alzheimer's disease.


Inclusion probability for DNA mixtures is a subjective one-sided match statistic unrelated to identification information.
DNA mixtures of two or more people are a common type of forensic crime scene evidence. A match statistic that connects the evidence to a criminal defendant is usually needed for court. Jurors rely on this strength of match to help decide guilt or innocence. However, the reliability of unsophisticated match statistics for DNA mixtures has been questioned. The most prevalent match statistic for DNA mixtures is the combined probability of inclusion (CPI), used by crime labs for over 15 years. When testing 13 short tandem repeat (STR) genetic loci, the CPI(-1) value is typically around a million, regardless of DNA mixture composition. However, actual identification information, as measured by a likelihood ratio (LR), spans a much broader range. This study examined probability of inclusion (PI) mixture statistics for 517 locus experiments drawn from 16 reported cases and compared them with LR locus information calculated independently on the same data. The log(PI(-1)) values were examined and compared with corresponding log(LR) values. The LR and CPI methods were compared in case examples of false inclusion, false exclusion, a homicide, and criminal justice outcomes. Statistical analysis of crime laboratory STR data shows that inclusion match statistics exhibit a truncated normal distribution having zero center, with little correlation to actual identification information. By the law of large numbers (LLN), CPI(-1) increases with the number of tested genetic loci, regardless of DNA mixture composition or match information. These statistical findings explain why CPI is relatively constant, with implications for DNA policy, criminal justice, cost of crime, and crime prevention. Forensic crime laboratories have generated CPI statistics on hundreds of thousands of DNA mixture evidence items. However, this commonly used match statistic behaves like a random generator of inclusionary values, following the LLN rather than measuring identification information. A quantitative CPI number adds little meaningful information beyond the analyst's initial qualitative assessment that a person's DNA is included in a mixture. Statistical methods for reporting on DNA mixture evidence should be scientifically validated before they are relied upon by criminal justice.


In-situ quantitative analysis of a prostate-specific antigen (PSA) using a nanomechanical PZT cantilever.
We report on a novel technique of resonant frequency shift measurement based on a nanomechanical cantilever with a PZT actuating layer for label-free detection of a prostate-specific antigen (PSA) in a liquid environment. The nanomechanical PZT thin film cantilever is composed of SiO(2)/Ta/Pt/PZT/Pt/SiO(2) on a SiN(x) supporting layer for simultaneous self-exciting and sensing; it was fabricated using a standard MEMS (micro electromechanical system) process. The specific binding characteristics of the PSA antigen to its antibody, which is immobilized with Calixcrown self-assembled monolayers (SAMs) on a gold surface deposited on a cantilever, are determined to a high sensitivity. For the bioassay in a liquid environment, a liquid test cell with a 20 microl volume reaction chamber has been fabricated, using a bonding technique between poly(dimethyl siloxane) (PDMS) bilayers. An observed trend of resonant frequency change with respect to time could be explained by the binding kinetics due to the Langmuir isotherm and diffusion and by the effects of a small volume reaction chamber. In the saturated regimes, the resonant frequency of the cantilever increased with increase of the PSA concentration in the reaction chamber, showing that the trend of the resonance frequency change was similar to that of the fluorescence results. The saturated resonance frequency shift of the cantilever was proportional to the PSA antigen concentration of analyte solution.


N-butyl cyanoacrylate glue embolization of arterial networks to facilitate hepatic arterial skeletonization before radioembolization.
Avoidance of nontarget microsphere deposition via hepatoenteric anastomoses is essential to the safety of yttrium-90 radioembolization (RE). The hepatic hilar arterial network may remain partially patent after coil embolization of major arteries, resulting in persistent risk. We retrospectively reviewed cases where n-butyl cyanoacrylate (n-BCA) glue embolization was used to facilitate endovascular hepatic arterial skeletonization before RE. A total of 543 RE procedures performed between June 2004 and March 2012 were reviewed, and 10 were identified where n-BCA was used to embolize hepatoenteric anastomoses. Arterial anatomy, prior coil embolization, and technical details were recorded. Outcomes were reviewed to identify subsequent complications of n-BCA embolization or nontarget RE. The rate of complete technical success was 80 % and partial success 20 %, with one nontarget embolization complication resulting in a minor change in treatment plan. No evidence of gastrointestinal or biliary ischemia or infarction was identified, and no microsphere-related gastroduodenal ulcerations or other evidence of nontarget RE were seen. Median volume of n-BCA used was <0.1 ml. n-BCA glue embolization is useful to eliminate hepatoenteric networks that may result in nontarget RE, especially in those that persist after coil embolization of major vessels such as the gastroduodenal and right gastric arteries.


Side-effect profile of fluoxetine in comparison with other SSRIs, tricyclic and newer antidepressants: a meta-analysis of clinical trial data.
In the last ten years, SSRIs have increasingly replaced TCAs as comparators of newer antidepressants (ADs), because of their better tolerability profile. In particular, fluoxetine has become a reference drug for the treatment of depression, but the occurrence of individual side effects in depressed subjects treated with fluoxetine and each comparator AD have not been systematically investigated. This meta-analysis investigated the frequency of side effects induced by fluoxetine or alternative ADs and compared the occurrence of individual side effects in depressed subjects. All randomised clinical trials (RCTs) comparing fluoxetine with any other AD drug in patients with major depression were located by searching the Cochrane Collaboration Depression, Anxiety and Neurosis Controlled Trials Register and the Cochrane Controlled Trials Register. Two reviewers independently extracted information. Significantly less percentage of patients treated with fluoxetine experienced any side effects in comparison with TCAs (50.9 % vs 60.3 %, 29 RCTs; RR = 0.84, p = 0.003), but not in comparison with other SSRIs (59.4 % vs 59.3 %, 15 RCTs; RR = 1.00, p = 0.902). In addition, fluoxetine was better tolerated in comparison with TCAs and related ADs (RR 0.61, 95 % CI 0.52, 0.71), but not in comparison with other SSRIs. Regard to individual side effects, activating (insomnia, agitation, tremor and anxiety) and gastrointestinal adverse events (nausea, vomiting, diarrhoea, weight loss and anorexia) were significantly more frequent in fluoxetine-treated patients, whereas cholinergic side effects were significantly less frequent. Fluoxetine compared to other ADs had more activating and gastrointestinal adverse effects, which often require additional pharmacotherapy or other managements strategies, leading to discontinuation and non-compliance and increasing the costs. This information is relevant to base on evidence the prescription of ADs in everyday clinical practice.


Structure of Mesophotic Reef Fish Assemblages in the Northwestern Hawaiian Islands.
Mesophotic coral ecosystems (MCEs) support diverse communities of marine organisms with changes in community structure occurring along a depth gradient. In recent years, MCEs have gained attention due to their depths that provide protection from natural and anthropogenic stressors and their relative stability over evolutionary time periods, yet ecological structures of fish assemblages in MCEs remain largely un-documented. Here, we investigated composition and trophic structure of reef fish assemblages in the Northwestern Hawaiian Islands (NWHI) along a depth gradient from 1 to 67 m. The structure of reef fish assemblages as a whole showed a clear gradient from shallow to mesophotic depths. Fish assemblages at mesophotic depths had higher total densities than those in shallower waters, and were characterized by relatively high densities of planktivores and invertivores and relatively low densities of herbivores. Fishes that typified assemblages at mesophotic depths included six species that are endemic to the Hawaiian Islands. The present study showed that mesophotic reefs in the NWHI support unique assemblages of fish that are characterized by high endemism and relatively high densities of planktivores. Our findings underscore the ecological importance of these undersurveyed ecosystems and warrant further studies of MCEs.


Genetic counseling in hereditary nonpolyposis colorectal cancer: an extended family with MSH2 mutation.
Molecular genetic advances have increased the demand for DNA testing. We describe DNA based genetic counseling in a hereditary nonpolyposis colorectal cancer (HNPCC) family. This extended HNPCC family was found to harbor the MSH2 germline mutation. Family history, medical, and pathology documents enabled us to secure a high degree of verification that the kindred qualified as HNPCC. DNA testing revealed the MSH2 germline mutation that was verified independently in two laboratories. Genetic counseling was provided before DNA testing and disclosure of MSH2 findings. Genetic counseling revealed a variety of findings characterized by emotional stress in MSH2 germline mutation carriers. Concerns centered around reproductive issues, potential transmission of the deleterious gene to their progeny, and discrimination by insurance carriers and employers. More than one-half of the patients found to harbor the MSH2 mutation considered the option of prophylactic subtotal colectomy. DNA testing should be restricted to well-verified candidate families in which genetic counseling should be mandatory. HNPCC family members sought genetic risk assessment for their own health and that of their children. Contrasting emotional responses took place when told of their gene testing status and this required a sensitive empathetic listening ear. Patients have many concerns about their lifetime cancer destiny when told that they harbor the culprit MSH2 germline mutation.


Vitamin D Deficiency and Oxidative Stress in Type 2 Diabetic Population of India.
Vitamin D deficiency is common in metropolitan cities of India and worldwide due to faulty life style. The prevalence of vitamin D deficiency has tremendously increased in India despite the fact that it is located between 8.4° and 37.6° north latitude and majority of the Indian population exposed to ample sunlight throughout the year. From published data regarding vitamin D deficiency in India estimated prevalence of vitamin D deficiency is 70%-100% in the adult population. The main identified reason for vitamin D deficiency is the lack of appropriate sunlight exposure and no consumption (in case of vegetarian population) or less consumption of vitamin D containing diet (e.g. fish, cod liver oil, beef liver and eggs). Mild to moderate vitamin D deficiency is identified as a risk factor for type 2 diabetes, due to its protective role against development of type 2 diabetes mellitus and its complications. Vitamin D may affect glucose homeostasis through increasing insulin resistance and reducing insulin secretion from beta cells of the pancreas. Vitamin D plays an important role against oxidative stress, because of its antioxidant property. Till date very few studies determined the link between vitamin D and oxidative stress in type 2 diabetes mellitus and its complications. However, more studies are needed to evaluate an underlying mechanism of vitamin D in amelioration of the oxidative stress in type 2 diabetes mellitus.


Implication of Ventricular Assist Devices in Extracorporeal Membranous Oxygenation Patients Listed for Heart Transplantation.
The new allocation criteria classify patients on veno-arterial extracorporeal membranous oxygenation (VA-ECMO) as the highest priority for receiving orthotopic heart transplantation (OHT) especially if they are considered not candidates for ventricular assist devices. The outcomes of patients who receive ventricular assist devices (VADs) after being listed for heart transplantation with VA-ECMO is unknown. We analyzed 355 patients listed for OHT with VA-ECMO from the United Network for Organ Sharing database from 2006 to 2014. Univariate and multivariate Cox proportional-hazards models were used to determine the contribution of prognostic variables to the outcome. Thirty-three patients (9.3%) received VADs (15 dischargeable, 7 non-dischargeable VADs). The VAD and non-VAD groups had similar listing characteristics except that the VAD group were more likely to have non-ischemic cardiomyopathy (48.5% vs. 25.2%), and less likely to be obese (6.1% vs. 25.2%) or have a history of prior organ transplant (3% vs. 31.1%). Patients who underwent VAD implantation had more days on the list (median 189 vs. 14 days) compared to the non-VAD group. Amongst the patients who had VADs, (25/33) 75.5% patients were subsequently transplanted with similar post-transplant survival compared to the non-VAD group (72% vs. 60.5%; p = 0.276). Predictors of one-year post-transplant mortality included panel reactive antibodies (PRA) class I ≥ 20%, recipient smoking history, increased serum creatinine and total bilirubin. Therefore, a small proportion of patients listed for transplantation with VA ECMO undergo VAD implantation. Their waitlist survival is better than non-VAD group but with similar post-transplant survival.


A comparison of the validity of GHQ-12 and CHQ-12 in Chinese primary care patients in Manchester.
The present study compares the efficacy of the GHQ-12 and the Chinese Health Questionnaire (CHQ-12) in Cantonese speaking Chinese primary-care patients living in Greater Manchester, using relative operating characteristic (ROC) analysis. We did not find that the Chinese version offered any advantage over the conventional version of the GHQ in this population. Stepwise discriminant analysis however confirmed the value of individual items in the former pertaining to specific somatic symptoms and interpersonal relationships in differentiating cases from non-cases. Information biases, arising from the lack of a reliability study on the second-stage case identifying interview and the unique linguistic characteristics of the Chinese language may have affected the overall validity indices of the questionnaires. The study also examines the effects of using different criteria to define a case, and shows that with increasing levels of severity, there is an improvement in the diagnostic performance of the two questionnaires as reflected by areas under ROC curves and traditional validity indices. Possible explanations of these findings are discussed. The scoring method proposed by Goodchild & Duncan-Jones (1985) when used on these questionnaires had no demonstrable advantage over the conventional scoring method.


The effects of 8 months of metformin on circulating GGT and ALT levels in obese women with polycystic ovarian syndrome.
The prevalence of non-alcoholic fatty liver disease (NAFLD) in polycystic ovarian syndrome (PCOS) is high. Small studies have shown reductions in serum alanine aminotransaminase (ALT) and gamma-glutamyltransaminase (GGT) concentrations, both surrogate liver fat markers, and sometimes improvements in liver histology in individuals with NAFLD treated with metformin. We investigated whether metformin reduces serum ALT and GGT concentrations in obese women with PCOS. We performed post hoc data analysis from a trial, involving 82 obese women aged 22-46 years with PCOS, conducted at an academic institution. Participants were treated with metformin 1500 or 2550 mg/day for 8 months. Anthropometric measurements and blood samples (serum ALT, GGT, lipids, leptin, C-reactive protein, insulin, glucose analyses) were taken at baseline, 4 and 8 months. Sixty-six participants completed the study. Mean weight, serum ALT and GGT decreased from 100.3 to 96.6 kg (p < 0.0001), 29.7 to 25.8 U/l (p = 0.012) and 21.4 to 16.9 U/l (p < 0.0001) respectively. Associations between weight reduction and decreases in serum ALT and GGT were highly significant and independent of change in Homeostasis Model Assessment of Insulin Resistance. In women with baseline ALT > 29.7 U/l (median), ALT reduction was highly significant (p = 0.005); however in those with baseline ALT < 29.7 U/l, ALT did not change despite similar weight reduction. There was no difference in reductions in ALT and GGT when the two metformin doses were compared. Intention-to-treat analyses gave comparable results. Metformin therapy is associated with reductions in surrogate liver fat markers in obese women with PCOS. This adds indirect support for a benefit of metformin in attenuating/reversing liver fat accumulation in PCOS and more generally.


Centers for Disease Control and Prevention Guideline for the Prevention of Surgical Site Infection, 2017.
The human and financial costs of treating surgical site infections (SSIs) are increasing. The number of surgical procedures performed in the United States continues to rise, and surgical patients are initially seen with increasingly complex comorbidities. It is estimated that approximately half of SSIs are deemed preventable using evidence-based strategies. To provide new and updated evidence-based recommendations for the prevention of SSI. A targeted systematic review of the literature was conducted in MEDLINE, EMBASE, CINAHL, and the Cochrane Library from 1998 through April 2014. A modified Grading of Recommendations, Assessment, Development, and Evaluation (GRADE) approach was used to assess the quality of evidence and the strength of the resulting recommendation and to provide explicit links between them. Of 5759 titles and abstracts screened, 896 underwent full-text review by 2 independent reviewers. After exclusions, 170 studies were extracted into evidence tables, appraised, and synthesized. Before surgery, patients should shower or bathe (full body) with soap (antimicrobial or nonantimicrobial) or an antiseptic agent on at least the night before the operative day. Antimicrobial prophylaxis should be administered only when indicated based on published clinical practice guidelines and timed such that a bactericidal concentration of the agents is established in the serum and tissues when the incision is made. In cesarean section procedures, antimicrobial prophylaxis should be administered before skin incision. Skin preparation in the operating room should be performed using an alcohol-based agent unless contraindicated. For clean and clean-contaminated procedures, additional prophylactic antimicrobial agent doses should not be administered after the surgical incision is closed in the operating room, even in the presence of a drain. Topical antimicrobial agents should not be applied to the surgical incision. During surgery, glycemic control should be implemented using blood glucose target levels less than 200 mg/dL, and normothermia should be maintained in all patients. Increased fraction of inspired oxygen should be administered during surgery and after extubation in the immediate postoperative period for patients with normal pulmonary function undergoing general anesthesia with endotracheal intubation. Transfusion of blood products should not be withheld from surgical patients as a means to prevent SSI. This guideline is intended to provide new and updated evidence-based recommendations for the prevention of SSI and should be incorporated into comprehensive surgical quality improvement programs to improve patient safety.


An RCOR1 loss-associated gene expression signature identifies a prognostically significant DLBCL subgroup.
Effective treatment of diffuse large B-cell lymphoma (DLBCL) is plagued by heterogeneous responses to standard therapy, and molecular mechanisms underlying unfavorable outcomes in lymphoma patients remain elusive. Here, we profiled 148 genomes with 91 matching transcriptomes in a DLBCL cohort treated with rituximab plus cyclophosphamide, doxorubicin, vincristine, and prednisolone (R-CHOP) to uncover molecular subgroups linked to treatment failure. Systematic integration of high-resolution genotyping arrays and RNA sequencing data revealed novel deletions in RCOR1 to be associated with unfavorable progression-free survival (P = .001). Integration of expression data from the clinical samples with data from RCOR1 knockdowns in the lymphoma cell lines KM-H2 and Raji yielded an RCOR1 loss-associated gene signature comprising 233 genes. This signature identified a subgroup of patients with unfavorable overall survival (P = .023). The prognostic significance of the 233-gene signature for overall survival was reproduced in an independent cohort comprising 195 R-CHOP-treated patients (P = .039). Additionally, we discovered that within the International Prognostic Index low-risk group, the gene signature provides additional prognostic value that was independent of the cell-of-origin phenotype. We present a novel and reproducible molecular subgroup of DLBCL that impacts risk-stratification of R-CHOP-treated DLBCL patients and reveals a possible new avenue for therapeutic intervention strategies.


Three-dimensional structure at the base of the mantle beneath the central pacific
Forward modeling of differential travel times of phases sensitive to lowermost mantle beneath the central Pacific reveals lateral heterogeneity that is higher in amplitude than predicted by tomographic models. A broad zone of low S velocity (-4 percent with respect to standard models), which may correspond to the base of a thermal "plume," narrows and is deflected as it extends to about 1000 kilometers above the core-mantle boundary. To the east of this zone, a localized region of fast S velocity (+5 percent) suggests strong heterogeneity or anisotropy related to the presence of high pressure and temperature assemblages, which may or may not involve core material. Its presence could also explain the observation of precursors to core reflected phases in this region.


A dodecanuclear copper(II) cage self-assembled from six dicopper building units.
Reaction of the dinucleating phenol-based ligand, H3bpmp (2,6-bis-[(3-hydroxy-propylimino)-methyl]-4-methyl-phenol), with Cu(2+) ions in the presence of a hybrid base (NEt3 and NaN3) necessary for the in situ generation of required numbers of hydroxido ions, results in the formation of a novel NO3(-) capped and HO(-) supported {Cu12} coordination complex {Cu6(μ3-OH)3(μ3-Hbpmp)3(μ3-NO3)}2(NO3)2(OH)2·2H2O·2MeOH (1). When the components are combined in right proportions (metal : ligand : NEt3 : NaN3 = 2 : 1 : 3 : 2) in MeOH, twelve Cu(2+) ions assemble in a cuboctahedral geometry, containing six square and eight triangular faces around a considerable void space. Six of the eight [Cu3] triangular faces are bound by the six Hbpmp(2-) ligands with six free pendant propanol arms around the central hexagonal plane. X-ray structure determination indicates new geometrical features for the core formation and reveals the face-capping potential of the H3bpmp ligand for the growth of a cuboctahedral coordination cage with the support of anions like HO(-) and NO3(-). The experimentally observed (J/kB = -173 K) strong antiferromagnetic coupling within the Cu12 complex has been justified by the DFT calculations.


Time-Resolved Systems Medicine Reveals Viral Infection-Modulating Host Targets.
Introduction: Drug-resistant infections are becoming increasingly frequent worldwide, causing hundreds of thousands of deaths annually. This is partly due to the very limited set of protein drug targets known for human-infecting viral genomes. The eleven influenza virus proteins, for instance, exploit host cell factors for replication and suppression of the antiviral immune responses. A systems medicine approach to identify relevant and druggable host factors would dramatically expand therapeutic options. Therapeutic target identification, however, has hitherto relied on static molecular networks, whereas in reality the interactome, in particular during an infection, is subject to constant change. Methods: We developed time-course network enrichment (TiCoNE), an expert-centered approach for discovering temporal response pathways. In the first stage of TiCoNE, time-series expression data is clustered in a human-augmented manner to identify groups of biological entities with coherent temporal responses. Throughout this process, the expert can add, remove, merge, or split temporal patterns. The resulting groups can then be mapped to an interaction network to identify enriched pathways and to analyze cross-talk enrichments and depletions between groups. Finally, temporal response groups of two experiments can be intersected, to identify condition-variant response patterns that represent promising drug-target candidates. Results: We applied TiCoNE to human gene expression data for influenza A virus infection and rhino virus infection, respectively. We then identified coherent temporal response patterns and employed our cross-talk analysis to establish two potential timelines of systems-level host responses for either infection. Next, we compared the two phenotypes and unraveled condition-variant temporal groups interacting on a networks level. The highest-ranking ones we then validated via literature search and wet-lab experiments. This not only confirmed many of our candidates as previously known, but we also identified phospholipid scramblase 1 (encoded by PLSCR1) as a previously not recognized host factor that is essential for influenza A virus infection. Conclusion: With TiCoNE we developed a novel approach for conjointly analyzing molecular networks with time-series expression data and demonstrated its power by identifying temporal drug-targets. We provide proof-of-concept that not only novel targets can be identified using our approach, but also that anti-infective drug target discovery can be enhanced by investigating temporal molecular networks of the host in response to viral infection.


The effect of human platelet lysate on corneal nerve regeneration.
This study aimed to test whether human platelet lysate (HPL) has neurotrophic ability for corneal nerve regeneration. We measured the neurotrophic factors in human peripheral serum (HPS) and two commercially available HPLs, UltraGRO and PLTMax. In vitro, we compared the growth rates, neuronal differentiation and immunostaining of neuron markers in mouse neuroblastoma cell line (Neuro-2a) and primary culture of mouse trigeminal ganglion cells that were cultivated in different concentrations of fetal bovine serum, HPS and HPL. In vivo, we created corneal wounds on Sprague Dawley rats with a rotating burr and evaluated the effects of topical HPL on wound healing and corneal nerve regeneration by in vivo confocal microscopy and corneal aesthesiometry. HPLs had significantly higher concentrations of various neurotrophic factors compared with HPS (p<0.05). In Neuro-2a cells, 3% HPL was better at promoting neuronal growth and differentiation compared with HPS at the same concentration. HPL was also found to have superior neurotrophic effects compared with HPS in primary cultures of mouse trigeminal ganglion cells. In vivo, HPL-treated eyes had better corneal epithelial wound healing rate, nerve regeneration length and corneal touch threshold compared with eyes treated with artificial tears (p<0.05). HPL has significantly higher concentrations of neurotrophic factors compared with HPS. It showed not only in vitro but also in vivo corneal neurotrophic abilities. Our results suggest that HPL may have a potential role in the treatment of diseases related to corneal nerve damage or degeneration.


Subclinical vitamin D deficiency in postmenopausal women with low vertebral bone mass.
To define the potential role of subclinical vitamin D deficiency in postmenopausal bone loss, we analyzed the levels of circulating 25-hydroxyvitamin D (25OHD) in 539 midwestern caucasian women screened for osteoporosis. Low 25OHD (less than 38 nmol/L) was found in 49 subjects (aged 52-77 yr). Women with low 25OHD had a reduced vertebral bone density (VBD), assessed by quantitative computed tomography, compared to age-matched controls (P less than 0.001). They also had significantly lower levels of serum calcium and phosphate, lower urinary calcium, higher serum alkaline phosphatase, and, in most cases, increased immunoreactive PTH (iPTH) concentrations, suggesting secondary hyperparathyroidism. Furthermore, only in the low 25OHD group did VBD correlate directly with 25OHD (r = 0.41; P less than 0.01), and inversely with iPTH (r = -0.47; P less than 0.01). Multivariate analyses revealed that iPTH was the major determinant of the observed decrease in VBD. Seasonal variations of serum 25OHD were noted only in the control population; in this group the 25OHD levels also correlated with sunlight exposure (r = 0.48; P less than 0.01), as assessed by an outdoor score. Thus, vitamin D deficiency develops when both the endogenous and exogenous sources are insufficient and contributes to a reduced bone mass in elderly women.


Play Behaviors in Children during the COVID-19 Pandemic: A Review of the Literature.
Play is a key factor for children's healthy psychological, emotional, social, and cognitive development. During the COVID-19 pandemic, it has been postulated that children's play was affected, not only regarding the time children spent playing but also in terms of the qualitative characteristics of play. The aim of this review was to investigate how children's play has changed during the COVID-19 pandemic. A review was conducted in the PubMed, Google Scholar, EMBASE, SCOPUS, ERIC, PsycInfo, and JSTOR databases up to 6 December 2020. Furthermore, references of eligible studies as well as of relevant articles were searched using a snowballing technique. The search retrieved 17 eligible studies, conducted in Europe and North America. In general, outdoor play was reduced during the pandemic; on the other hand, there was an increase in indoor play and in videogames-screen time. COVID-19 was present in children's pretend play. Children's play was a key contributor to children's mood and wellbeing. Furthermore, teachers were especially concerned about how children's play was affected during the lockdown measures. There is evidence that children's play habits were affected during the COVID-19 pandemic; further research is required, especially cross-culturally oriented.


Coadministration of seasonal influenza vaccine and MVA-NP+M1 simultaneously achieves potent humoral and cell-mediated responses.
Current seasonal influenza vaccines have reduced immunogenicity and are of suboptimal efficacy in older adults. We have previously shown that the novel candidate vaccine MVA-NP+M1 is able to boost memory T cell responses in adults aged 50-85 years. Preclinical studies have demonstrated that viral vectored vaccines can act as adjuvants when coadministered with protein-based vaccines. We have conducted a phase I clinical trial to compare the coadministration of seasonal influenza vaccine and MVA-NP+M1 with seasonal influenza vaccine alone in adults aged 50 years and above. This combination of vaccines was safe and well tolerated. T cell responses to internal influenza proteins were boosted to significantly higher levels in the group receiving MVA-NP+M1 compared with the group receiving seasonal influenza vaccine alone. Rates of seroprotection and seroconversion against the three vaccine strains were similar in both groups; however, there was a significant increase in the geometric mean titer ratio for the H3N2 component of seasonal influenza vaccine in the coadministration group. While some vaccine combinations result in immune interference, the coadministration of MVA-NP+M1 alongside seasonal influenza vaccine is shown here to increase some influenza strain-specific antibody responses and boost memory T cells capable of recognizing a range of influenza A subtypes.


[LEVELS OF OBESITY, METABOLIC PROFILE, CONSUMPTION OF TABACO AND BLOOD PRESSURE IN SEDENTARY YOUTHS].
in Chile, the National Health Survey (ENS) conducted in 2009-2010 reported high prevalence of overweight, sedentary lifestyle, high cholesterol and metabolic syndrome in the population. to determine the prevalence in young sedentary obesity and consumption of tabaco and analyze their association with the metabolic profile, body fat percentage and blood pressure. 125 young sedentary, 26 men and 99 women, aged between 17 and 29 years old were evaluated. Body mass index (BMI), percent body fat (% fat), waist contour (CC), systolic and diastolic blood pressure, total cholesterol, HDL cholesterol (HDL-C), LDL cholesterol (LDL-C), triglycerides: measurements were performed, glycemia and consumption of snuff. HDL-C (p = 0.000) and% MG (p = 0.043) were higher in women. 37.6% of young people turned smoker. 35, 2% of the sample showed excessive malnutrition. Obese subjects had higher levels: waist contour (p = 0.000) and% FM (p = 0.000). When analyzing obesity DC, this showed significant differences in BMI,% fat, systolic and diastolic blood pressure. BMI presented positive association with CC,% fat, total cholesterol, triglycerides, LDL, systolic and diastolic blood pressure (p < 0.05). The CC presented association with MG%, triglycerides, systolic blood pressure and diastolic (p < 0.05). The MG% did not present association with other variables a high prevalence of malnutrition by excess consumption of tabaco in the study sample, while other variables are not high-risk categories, it is an opportune time to intervene and reverse these negative health trends now.


Intravitreous ranibizumab for persistent macular edema in retinal vein occlusion unresponsive to bevacizumab.
To report successful treatment of persistent retinal edema associated with retinal vascular occlusion with ranibizumab after initial failure with bevacizumab intravitreal injections. A retrospective chart review of all patients undergoing treatment with intravitreal bevacizumab for retinal vascular occlusion was undertaken. Clinical features of patients who were switched to monthly ranibizumab because of persistent and/or worsening retinal edema despite multiple (six or more) monthly intravitreal bevacizumab injections were selected for review. The type of retinal vascular occlusion, age of the patient, visual acuity at each visit, number of intravitreal bevacizumab injections, optical coherence tomography findings at each visit, fluorescein angiography, and number of intravitreal ranibizumab injections with optical coherence tomography findings at each subsequent visit were recorded. Six patients (five women and one man) were identified. Age ranged from 56 years to 74 years. Five had central retinal vein occlusion and one a branch retinal vein occlusion. All were treated with at least 6 sequential intravitreal bevacizumab injections (range, 6-13) before treatment with ranibizumab. All demonstrated an immediate and marked reduction in retinal edema that had not occurred with any of the previous treatments. The average central foveal thickness on optical coherence tomography was 592 µm before ranibizumab injection and 346 µm after the injection. Visual acuity improved in four of the six patients and was stable in the remaining two patients. Eyes of patients with persistent macular edema secondary to retinal vein occlusion despite bevacizumab may experience prompt improvement after initiation of intravitreal ranibizumab injections.