Wound healing/regeneration using recombinant human growth/differentiation factor-5 in an injectable poly-lactide-co-glycolide-acid composite carrier and a one-wall intra-bony defect model in dogs.
The purpose of this study was to evaluate the effect of recombinant human growth/differentiation factor-5 (rhGDF-5) on periodontal wound healing/regeneration using an injectable poly-lactide-co-glycolide-acid (PLGA) composite carrier and an established defect model. Bilateral 4 × 5 mm (width × depth) one-wall, critical-size, intra-bony periodontal defects were surgically created at the second and the fourth mandibular pre-molar teeth in 15 Beagle dogs. The animals were randomized to receive (using a split-mouth design; defect sites in the same jaw quadrant getting the same treatment) rhGDF-5 high dose (188 μg/defect) versus sham-surgery control (five animals), rhGDF-5 mid dose (37 μg/defect) versus carrier control (five animals), and rhGDF-5 low dose (1.8 μg/defect) versus treatment reported elsewhere (five animals). The animals were euthanized for histometric analysis following an 8-week healing interval. Clinical healing was uneventful. The rhGDF-5/PLGA construct was easy to assemble and apply. The rhGDF-5 high dose supported significantly increased bone formation compared with the low-dose, sham-surgery, and carrier controls (p<0.05) and induced significantly increased cementum formation compared with the controls (p<0.05). Root resorption/ankylosis or other aberrant healing events were not observed. rhGDF-5 appears to effectively support periodontal wound healing/regeneration in a dose-dependent order; the PLGA composite appears to be an effective ease-of-use candidate for carrier technology.


Synthesis, structural characterisation, and synthetic application of stable seleniranium ions.
Stable seleniranium ions were prepared from easily available stable bromiranium ions and diselenides. The solid state structure of the obtained seleniranium ions was determined by X-ray crystallographic analysis and their alkene-to-alkene transfer was investigated by NMR techniques. The rapid alkene-to-alkene transfer of the selenium group enabled the application of the seleniranium ion salts as selenenylating agents, which led to very efficient and highly diastereoselective, selenium-induced polyene-type cyclisations of terpene analogues.


Stable ultra-thin CdTe crystal: a robust direct gap semiconductor.
Employing density functional theory based calculations, we investigate structural, vibrational and strain-dependent electronic properties of an ultra-thin CdTe crystal structure that can be derived from its bulk counterpart. It is found that this ultra-thin crystal has an 8-atom primitive unit cell with considerable surface reconstructions. Dynamic stability of the structure is predicted based on its calculated vibrational spectrum. Electronic band structure calculations reveal that both electrons and holes in single layer CdTe possess anisotropic in-plane masses and mobilities. Moreover, we show that the ultra-thin CdTe has some interesting electromechanical features, such as strain-dependent anisotropic variation of the band gap value, and its rapid increase under perpendicular compression. The direct band gap semiconducting nature of the ultra-thin CdTe crystal remains unchanged under all types of applied strain. With a robust and moderate direct band gap, single-layer CdTe is a promising material for nanoscale strain dependent device applications.


Input-dependent synaptic targeting of alpha(2)-subunit-containing GABA(A) receptors in synapses of hippocampal pyramidal cells of the rat.
Pyramidal cells, expressing at least 14 subunits of the heteropentameric GABA(A) receptor, receive GABAergic input on their soma and proximal dendrites from basket cells, activating GABA(A) receptors and containing either parvalbumin or cholecystokinin and vasoactive intestinal polypeptide. The properties of GABA(A) receptors are determined by the subunit composition, and synaptic receptor content governs the effect of the presynaptic neuron. Using a quantitative electron microscopic immunogold technique, we tested whether the synapses formed by the two types of basket cell show a difference in the subunit composition of GABA(A) receptors. Terminals of one of the basket cells were identified by antibodies to parvalbumin. Synapses made by parvalbumin-negative terminals showed five times more immunoreactivity for the alpha(2) subunit than synapses made by parvalbumin-positive basket cells, whose synapses were frequently immunonegative. This difference is likely to be due to specific GABA(A) receptor alpha subunit composition, because neither synaptic size nor immunoreactivity for the beta(2/3) subunits, indicating total receptor content, was different in these two synapse populations. Synapses established by axo-axonic cells on axon initial segments showed an intermediate number of immunoparticles for the alpha(2) subunit compared to those made by basket cells but, due to their smaller size, the density of the alpha(2) subunit immunoreactivity was higher in synapses on the axon. Because the two basket cell types innervate the same domain of the pyramidal cell, the results indicate that pyramidal cells have mechanisms to target GABA(A) receptors, under presynaptic influence, preferentially to distinct synapses. The two basket cell types act via partially distinct GABA(A) receptor populations.


A double-blind comparison of sertraline and fluoxetine in depressed elderly outpatients.
There has been a paucity of well-designed studies comparing selective serotonin reuptake inhibitor (SSRI) medications in the treatment of depression in the elderly. This multicenter study was designed to examine the efficacy and safety of sertraline and fluoxetine in depressed elderly outpatients. A secondary objective was to examine the effects of SSRI treatment on quality of life and cognitive function. Two hundred thirty-six outpatients 60 years of age and older who met DSM-III-R criteria for major depressive disorder received 1 week of single-blind placebo before being randomly assigned to 12 weeks of double-blind, parallel-group treatment with flexible daily doses of either sertraline (range, 50-100 mg) or fluoxetine (range, 20-40 mg). Primary efficacy measures consisted of the 24-item Hamilton Rating Scale for Depression and Clinical Global Impressions scale ratings. Secondary outcome assessments included clinician- and patient-rated measures of depression symptoms and factors, cognitive functioning, and quality of life, as well as plasma drug concentrations, which were correlated with clinical response. Both drugs produced a similarly positive response on the primary efficacy measures, with 12-week responder rates of 73% for sertraline and 71% for fluoxetine. Sertraline-treated patients showed statistically greater cognitive improvement on several measures. Both drugs were safe and well tolerated. Data indicate that both drugs are effective antidepressants for the treatment of depressed elderly outpatients. Differences in cognitive performance effects deserve further investigation.


Nocturnal Hypertension: Neglected Issue in Comprehensive Hypertension Management.
The body circardian rhythm affects blood pressure variability at day and night, therefore blood pressure at day and night might be different. Nocturnal hypertension is defined as increase of blood pressure >120/70mmHg at night, which is caused by disturbed circadian rhythm, and associated with higher cardiovascular and cerebrovascular events also mortality in hypertensive patients. Nocturnal hypertension and declining blood pressure pattern, can only be detected by continuous examination for 24 hours, also known as ambulatory blood pressure measurement (ABPM). Chronotherapy, has become a strategy for managing the hypertensive nocturnal patients, by taking hypertensive medication at night to obtain normal blood pressure decrease in accordance with the normal circadian rhythm and, improving blood pressure control.


Patients' self-reported adherence to cardiovascular medication using electronic monitors as comparators.
The aim of this study was to evaluate self-reporting of adherence to cardiovascular medication using electronic pillboxes (medical event monitoring system [MEMS]) as the gold standard comparator. In total, 78 individuals (52% hypertensives, 21% diabetics, 27% with dyslipidemia) were recruited prospectively from an outpatient clinic setting in Switzerland. Participants completed two self-report measures (visual analogue scale [VAS] and a validated self-reporting questionnaire) at baseline and were asked to use MEMS as their pillbox for the subsequent 10 weeks. Patients expressed their medication adherence behaviour on a VAS (0 mm="I never take any tablets"; 100 mm="I take all tablets as prescribed") and entered one of six numbers (from 1: perfect adherence to 6: non-adherence) on the questionnaire. Medication compliance was monitored for 75 d on average. Mean (+/-SD, range) scores for MEMS with respect to timing adherence, correct dosing, and self-administration adherence were 79+/-25% (8-100%), 83+/-20% (24-100%), and 92+/-17% (54-118%), respectively. A majority of participants (78.8%) over-reported their adherence to the VAS (93+/-7 mm, 73-100), and VAS scores correlated poorly with MEMS recordings (Spearman's rho for timing adherence, correct dosing, self-administration adherence 0.29 [p=0.018], 0.24 [p=0.051], 0.26 [p=0.036], respectively). Similarly, we found no correlation between adherence as expressed in the questionnaire and MEMS (regression coefficients <0.1). We conclude that a majority of patients over-report adherence to cardiovascular medication if asked to complete a visual analogue scale and a validated questionnaire. Therefore, using self-reporting as the sole means of assessing medication compliance is insufficiently accurate to detect poor adherence. (Hypertens Res 2008; 31: 2037-2043).


Mediterranean diet reduces 24-hour ambulatory blood pressure, blood glucose, and lipids: one-year randomized, clinical trial.
The PREvención con DIeta MEDiterránea (PREDIMED) trial showed that Mediterranean diets (MedDiets) supplemented with either extravirgin olive oil or nuts reduced cardiovascular events, particularly stroke, compared with a control, lower fat diet. The mechanisms of cardiovascular protection remain unclear. We evaluated the 1-year effects of supplemented MedDiets on 24-hour ambulatory blood pressure (BP), blood glucose, and lipids. Randomized, parallel-design, controlled trial was conducted in 2 PREDIMED sites. Diets were ad libitum, and no advice on increasing physical activity or reducing sodium intake was given. Participants were 235 subjects (56.5% women; mean age, 66.5 years) at high cardiovascular risk (85.4% with hypertension). Adjusted changes from baseline in mean systolic BP were -2.3 (95% confidence interval [CI], -4.0 to -0.5) mm Hg and -2.6 (95% CI, -4.3 to -0.9) mm Hg in the MedDiets with olive oil and the MedDiets with nuts, respectively, and 1.7 (95% CI, -0.1 to 3.5) mm Hg in the control group (P<0.001). Respective changes in mean diastolic BP were -1.2 (95% CI, -2.2 to -0.2), -1.2 (95% CI, -2.2 to -0.2), and 0.7 (95% CI, -0.4 to 1.7) mm Hg (P=0.017). Daytime and nighttime BP followed similar patterns. Mean changes from baseline in fasting blood glucose were -6.1, -4.6, and 3.5 mg/dL (P=0.016) in the MedDiets with olive oil, MedDiets with nuts, and control diet, respectively; those of total cholesterol were -11.3, -13.6, and -4.4 mg/dL (P=0.043), respectively. In high-risk individuals, most with treated hypertension, MedDiets supplemented with extravirgin olive oil or nuts reduced 24-hour ambulatory BP, total cholesterol, and fasting glucose. http://www.clinicaltrials.gov. Unique identifier: ISRCTN35739639.


Reflection of focused beams from opal photonic crystals.
We present a robust method for computing the reflection of arbitrarily shaped and sized beams from finite thickness photonic crystals. The method is based on dividing the incident beam into plane waves, each of which can be solved individually using Bloch periodic boundary conditions. This procedure allows us to take a full advantage of the crystal symmetry and also leads to a linear scaling of the computation time with respect to the number of plane waves needed to expand the incident beam. The algorithm for computing the reflection of an individual plane wave is also reviewed. Finally, we find an excellent agreement between the computational results and measurement data obtained from opals that are synthesized using polystyrene and poly(methyl methacrylate) microspheres.


Ethical dilemmas among dental professionals in Davangere city- a cross-sectional survey.
To determine the ethical problems faced by dental practitioners, a cross-sectional survey was conducted in Bapuji Dental College and Hospital, College of Dental Sciences and private dental clinics in the city of Davangere. A questionnaire with close-ended questions on eight scenarios was administered to the 135 study participants. In the case of scenario 1, 81.4% of the participants said that the doctor had violated the principle of truthfulness. As for scenario 2, less than 50% of the participants replied that the doctor had breached ethical principles. In case scenario 3, 93% felt that the doctor should have taken the physician's opinion before extracting a tooth. Most dental practitioners faced ethical dilemmas because of the lack of awareness, and there is a need to introduce certain programmes to promote knowledge of ethics.


Human diploid fibroblast cells in senescence; cycling through polyploidy to mitotic cells.
Previously, it was found that senescent cells can undergo a modified cell cycle with mitotic cells as the end results. The major cycling events started with polyploidization, followed by depolyploidization to multinucleated cells (MNCs). These latter cells produced mononuclear offspring cells that could express mitotic cell divisions. In this report the emphasis is on late senescent fibroblasts that exhibited the senescence-associated change in cell morphology to large flat cells. Prior to live cell photography, flat cell cultures were maintained for months in the same culture flasks and therefore judged to be in a late senescent phase. All of the cellular events outlined above were present in these old cell cultures. Time lapse pictures showed movements of mitotic daughter cells away from each other and alignment of the chromosomes on the metaphase plate was visible in other mitotic cells. These data challenge the common view that cell senescence is irreversible and, therefore, an antitumor mechanism. A new finding was that the spike in polyploid cells in the near senescent phase consisted of cells with pairs of sister chromosomes from endoreduplication of DNA (two rounds of DNA synthesis and no mitosis). The lack of cells with 92 single chromosomes (e.g., G2 tetraploid cells) suggested that these polyploid cells also went through a changed cell cycle. The question now is whether these atypical polyploid cells are a subpopulation in senescence that can undergo the cycling from polyploidy to genome-reduced mitotic cells.


Transapical aortic valve implantation: step by step.
Transapical aortic valve implantation is a new minimally invasive technique for beating heart, off-pump, aortic valve implantation in high-risk patients. The procedure involves antegrade aortic valve implantation using an oversizing technique with direct access and accurate positioning of a stent-based transcatheter xenograft. Procedural steps include placement of femoral arterial and venous access wires, anterolateral mini-thoracotomy, epicardial pacing, and apical pursestring suture placement. Valve positioning is performed under fluoroscopic and echocardiographic guidance during rapid ventricular pacing. Patient screening, especially regarding native aortic annulus diameter and pattern of calcification, is essential for success. Since imaging is crucial, implantations are optimally performed in a hybrid operative theater by an experienced team of cardiac surgeons, cardiologists, and anesthetists. The aim of this article is to outline the technical aspects of the new technique of minimally invasive transapical aortic valve implantation.


Incidental findings on brain MR imaging in older community-dwelling subjects are common but serious medical consequences are rare: a cohort study.
Incidental findings in neuroimaging occur in 3% of volunteers. Most data come from young subjects. Data on their occurrence in older subjects and their medical, lifestyle and financial consequences are lacking. We determined the prevalence and medical consequences of incidental findings found in community-dwelling older subjects on brain magnetic resonance imaging. Prospective cohort observational study. Single centre study with input from secondary care. Lothian Birth Cohort 1936, a study of cognitive ageing. Incidental findings identified by two consultant neuroradiologists on structural brain magnetic resonance imaging at age 73 years; resulting medical referrals and interventions. PREVALENCE OF INCIDENTAL FINDINGS BY INDIVIDUAL CATEGORIES: neoplasms, cysts, vascular lesions, developmental, ear, nose or throat anomalies, by intra- and extracranial location; visual rating of white matter hyperintensities and brain atrophy. There were 281 incidental findings in 223 (32%) of 700 subjects, including 14 intra- or extracranial neoplasms (2%), 15 intracranial vascular anomalies (2%), and 137 infarcts or haemorrhages (20%). Additionally, 153 had moderate/severe deep white matter hyperintensities (22%) and 176 had cerebral atrophy at, or above, the upper limit of normal (25%) compared with a normative population template. The incidental findings were unrelated to white matter hyperintensities or atrophy; about a third of subjects had both incidental findings and moderate or severe WMH and a quarter had incidental findings and atrophy. The incidental findings resulted in one urgent and nine non-urgent referrals for further medical assessment, but ultimately in no new treatments. In community-dwelling older subjects, incidental findings, including white matter hyperintensities and atrophy, were common. However, many findings were not of medical importance and, in this age group, most did not result in further assessment and none in change of treatment.


Replication of phage phi 29 DNA with purified terminal protein and DNA polymerase: synthesis of full-length phi 29 DNA.
A system that replicates bacteriophage phi 29 DNA with protein p3 covalently attached to the two 5' ends, using as the only proteins the phi 29 DNA polymerase and the terminal protein, is described. Restriction analysis of the 32P-labeled DNA synthesized in vitro showed that all phi 29 DNA fragments were labeled. Analysis by alkaline sucrose gradient centrifugation of the DNA labeled during a 10-min pulse showed that, after a 20-min chase, about half of the DNA molecules had reached apparently full-length phi 29 DNA (approximately equal to 18,000 nucleotides). Ammonium ions strongly stimulated phi 29 DNA-protein p3 replication, the effect being due to stimulation of the initiation reaction. ATP was not required for phi 29 DNA-protein p3 replication, either in the initiation or elongation steps. The results show that the phi 29 DNA polymerase functions, not only in the formation of the p3-dAMP covalent initiation complex but also in the elongation of the latter, as the only DNA polymerase to produce full-length phi 29 DNA.


Expression of insulin-like growth factor system components in Ewing's sarcoma and their association with survival.
The role of IGF system in the pathogenesis of Ewing's sarcoma (EWS) is well-documented. However, still little information is available about the value of IGF system components as indicators of prognosis. Understanding the clinical role for IGF system in EWS patients may be important because different subtypes of patients have distinct outcome and may require different treatment protocol. We evaluated the expression of insulin-like growth factor (IGF)-receptor (IGF-IR), insulin receptor (IR), IGF-I and some major intracellular mediators (IRS1, p-ERK) in specimens from EWS patients with primary localised untreated tumours. 290 samples were used for immunohistochemistry studies; 57 samples for Real-Time PCR studies; and serum of 67 patients for ELISA. IGF-IR and IR are expressed in virtually all EWS tumours. Usually both the receptors are present in the same tumour but when one receptor is lacking the other one is always present. Evaluation of IGF-IR, IR and IGF-I with quantitative methods may discriminate differential levels of expression with influences on the patients' outcome. High expression of IGF-IR, IR and IGF-I mRNAs is significantly associated with more favourable clinical outcomes. Higher circulating levels of IGF-I also correlated with lower risk to disease progression and death. Overall, our clinical data are in contrast with the assumption that higher amounts of IGF/IGF-IR are a surrogate for higher aggressiveness and indicate that in some cancers the transition to frank malignancy and poor treatment responsiveness seem to be associated with a reduction of IGF system activity.


Common variants underlying cognitive ability: further evidence for association between the SNAP-25 gene and cognition using a family-based study in two independent Dutch cohorts.
The synaptosomal associated protein of 25 kDa (SNAP-25) gene, located on chromosome 20 p12-12p11.2 encodes a presynaptic terminal protein. SNAP-25 is differentially expressed in the brain, and primarily present in the neocortex, hippocampus, anterior thalamic nuclei, substantia nigra and cerebellar granular cells. Recently, a family-based genetic association was reported between variation in intelligence quotient (IQ) phenotypes and two intronic variants on the SNAP-25 gene. The present study is a follow-up association study in two Dutch cohorts of 371 children (mean age 12.4 years) and 391 adults (mean age 36.2 years). It examines the complete genomic region of the SNAP-25 gene to narrow down the location of causative genetic variant underlying the association. Two new variants in intron 1 (rs363043 and rs353016), close to the two previous reported variants (rs363039 and rs363050) showed association with variation in IQ phenotypes across both cohorts. All four single nucleotide polymorphisms were located in intron 1, within a region of about 13.8 kbp, and are known to affect transcription factor-binding sites. Contrary to what is expected in monogenic traits, subtle changes are postulated to influence the phenotypic outcome of complex (common) traits. As a result, functional polymorphisms in (non)coding regulatory sequences may affect spatial and temporal regulation of gene expression underlying normal cognitive variation.


Independent predictors of morbidity after image-guided stereotactic brain biopsy: a risk assessment of 270 cases.
Image-guided stereotactic brain biopsy is associated with transient and permanent incidences of morbidity in 9 and 4.5% of patients, respectively. The goal of this study was to perform a critical analysis of risk factors predictive of an enhanced operative risk in frame-based and frameless stereotactic brain biopsy. The authors reviewed the clinical and neuroimaging records of 270 patients who underwent consecutive frame-based and frameless image-guided stereotactic brain biopsies. The association between preoperative variables and biopsy-related morbidity was assessed by performing a multivariate logistic regression analysis. Transient and permanent stereotactic biopsy-related morbidity was observed in 23 (9%) and 13 (5%) patients, respectively. A hematoma occurred at the biopsy site in 25 patients (9%); 10 patients (4%) were symptomatic. Diabetes mellitus (odds ratio [OR] 3.73, 95% confidence interval [CI] 1.37-10.17, p = 0.01), thalamic lesions (OR 4.06, 95% CI 1.63-10.11, p = 0.002), and basal ganglia lesions (OR 3.29, 95% CI 1.05-10.25, p = 0.04) were in'dependent risk factors for morbidity. In diabetic patients, a serum level of glucose that was greater than 200 mg/dl on the day of biopsy had a 100% positive predictive value and a glucose level lower than 200 mg/dl on the same day had a 95% negative predictive value for biopsy-related morbidity. Pontine biopsy was not a risk factor for morbidity. Only two (4%) of 45 patients who had epilepsy before the biopsy experienced seizures postoperatively. The creation of more than one needle trajectory increased the incidence of neurological deficits from 17 to 44% when associated with the treatment of deep lesions (those in the basal ganglia or thalamus; p = 0.05), but was not associated with morbidity when associated with the treatment of cortex lesions. Basal ganglia lesions, thalamic lesions, and patients with diabetes were independent risk factors for biopsy-associated morbidity. Hyperglycemia on the day of biopsy predicted morbidity in the diabetic population. Epilepsy did not predispose to biopsy-associated seizure. For deep-seated lesions, increasing the number of biopsy samples along an established track rather than performing a second trajectory may minimize the incidence of morbidity. Close perioperative observation of glucose levels may be warranted.


Increased intestinal epithelial proliferation in metallothioneine-transforming growth factor alpha transgenic mice.
The epidermal growth factor (EGF) peptide family includes six closely-related proteins, all of which bind to the EGF receptor. In the intestinal epithelium, transforming growth factor alpha (TGFalpha) appears to be the most physiological ligand for the EGF receptor. The present studies were designed to examine the effect of TGFalpha overexpression on duodenal epithelial proliferation using a metallothioneine-inducible promoter/enhancer transgenic mouse (MT-TGFalpha). The MT-TGFalpha mouse model was further studied to determine the in vivo effect of unregulated TGFalpha production on the physiological proliferative responses to fasting and refeeding. MT-TGFalpha mice were given 25 mM oral ZnSO4 to induce transgene expression and were studied 1 to 2 months later. Duodenal histology was analyzed morphometrically in well-oriented transverse sections. The vincristine metaphase-arrest technique was used to assess proliferation in duodenal crypts. Immunohistochemical staining and in situ hybridization were used to assess transgenic TGFalpha protein and mRNA expression, respectively. Normally fed MT-TGFalpha mice had deeper crypts (0.12 vs. 0.08 mm), longer villi (0.66 vs. 0.54 mm), and greater luminal diameters (2.65 vs. 2.19 mm) compared to controls (P<0.05 for all three dimensions). The crypt cell mitotic index in normally fed transgenic mice was 1.5 fold greater than the index in normally fed controls (20+/-2 vs. 35+/-4 mitoses per crypt; P <0.05). Fasting and refeeding MT-TGFalpha mice resulted in no significant change in their high baseline rate of crypt proliferation, while proliferation in control mice rose from a lower baseline during fasting to a level with refeeding that approximated rates in MT-TGFalpha mice. Transgenic TGFalpha protein and mRNA were localized to the villus epithelial compartment with little or no evidence of mRNA or protein expression in the crypt epithelium. Overproduction of TGFalpha in the mouse duodenal epithelium results in a pronounced increase in crypt epithelial cell proliferation and a resulting increase in the dimension of the crypt/villus unit. This appears to be mediated through a paracrine and/or juxtacrine effect on crypt cells by TGFalpha produced in the villus epithelium. Fasting and refeeding experiments suggest that TGFalpha may also play a role in the proliferative response to refeeding or that the full potential for proliferation is realized by TGFalpha overexpression alone. Collectively, these studies suggest that TGFalpha is a physiological autocrine and paracrine regulator of small intestinal epithelial proliferation.


Prenatal Depression and Infant Health: The Importance of Inadequately Measured, Unmeasured, and Unknown Confounds.
A recent study found that maternal antenatal depressive symptoms were associated with adverse infant general health outcomes and that gestational age, birth weight, and breastfeeding did not mediate the observed relationship. The authors suggested that antenatal depression can have a harmful effect on infant health through disturbed fetal programming driven by maternal symptoms and behaviors that influence the maternal and hence the fetal internal environment. The authors implied that interventions to diagnose and treat maternal depression can have a protective effect against disturbances in infant health. However, because of the observational nature of the study, cause-effect relationships cannot be conclusively stated. This is especially so because there were many confounds that the authors did not consider. The present article provides examples and explanations of how inadequately measured, unmeasured, and unknown confounds can explain observed relationships between explanatory and outcome variables, thereby negating cause-effect interpretations of study findings. It is important to minimize confounding when conducting observational studies, and this can only be done by comprehensively listing and efficiently measuring potential confounders in advance.


Antigenic and genetic variation in cytopathic hepatitis A virus variants arising during persistent infection: evidence for genetic recombination.
Variants of hepatitis A virus (pHM175 virus) recovered from persistently infected green monkey kidney (BS-C-1) cells induced a cytopathic effect during serial passage in BS-C-1 or fetal rhesus kidney (FRhK-4) cells. Epitope-specific radioimmunofocus assays showed that this virus comprised two virion populations, one with altered antigenicity including neutralization resistance to monoclonal antibody K24F2, and the other with normal antigenic characteristics. Replication of the antigenic variant was favored over that of virus with the normal antigenic phenotype during persistent infection, while virus with the normal antigenic phenotype was selected during serial passage. Viruses of each type were clonally isolated; both were cytopathic in cell cultures and displayed a rapid replication phenotype when compared with the noncytopathic passage 16 (p16) HM175 virus which was used to establish the original persistent infection. The two cytopathic virus clones contained 31 and 34 nucleotide changes from the sequence of p16 HM175. Both shared a common 5' sequence (bases 30 to 1677), as well as sequence identity in the P2-P3 region (bases 3249 to 5303 and 6462 to 6781) and 3' terminus (bases 7272 to 7478). VP3, VP1, and 3Cpro contained different mutations in the two virus clones, with amino acid substitutions at residues 70 of VP3 and 197 and 276 of VP1 of the antigenic variant. These capsid mutations did not affect virion thermal stability. A comparison of the nearly complete genomic sequences of three clonally isolated cytopathic variants was suggestive of genetic recombination between these viruses during persistent infection and indicated that mutations in both 5' and 3' nontranslated regions and in the nonstructural proteins 2A, 2B, 2C, 3A, and 3Dpol may be related to the cytopathic phenotype.


The Fungal Exopolysaccharide Galactosaminogalactan Mediates Virulence by Enhancing Resistance to Neutrophil Extracellular Traps.
Of the over 250 Aspergillus species, Aspergillus fumigatus accounts for up to 80% of invasive human infections. A. fumigatus produces galactosaminogalactan (GAG), an exopolysaccharide composed of galactose and N-acetyl-galactosamine (GalNAc) that mediates adherence and is required for full virulence. Less pathogenic Aspergillus species were found to produce GAG with a lower GalNAc content than A. fumigatus and expressed minimal amounts of cell wall-bound GAG. Increasing the GalNAc content of GAG of the minimally pathogenic A. nidulans, either through overexpression of the A. nidulans epimerase UgeB or by heterologous expression of the A. fumigatus epimerase Uge3 increased the amount of cell wall bound GAG, augmented adherence in vitro and enhanced virulence in corticosteroid-treated mice to levels similar to A. fumigatus. The enhanced virulence of the overexpression strain of A. nidulans was associated with increased resistance to NADPH oxidase-dependent neutrophil extracellular traps (NETs) in vitro, and was not observed in neutropenic mice or mice deficient in NADPH-oxidase that are unable to form NETs. Collectively, these data suggest that cell wall-bound GAG enhances virulence through mediating resistance to NETs.


Is evidence for homoeopathy reproducible?
We tested, under independent conditions, the reproducibility of evidence from two previous trials that homoeopathy differs from placebo. The test model was again homoeopathic immunotherapy. 28 patients with allergic asthma, most of them sensitive to house-dust mite, were randomly allocated to receive either oral homoeopathic immunotherapy to their principal allergen or identical placebo. The test treatments were given as a complement to their unaltered conventional care. A daily visual analogue scale of overall symptom intensity was the outcome measure. A difference in visual analogue score in favour of homoeopathic immunotherapy appeared within one week of starting treatment and persisted for up to 8 weeks (p = 0.003). There were similar trends in respiratory function and bronchial reactivity tests. A meta-analysis of all three trials strengthened the evidence that homoeopathy does more than placebo (p = 0.0004). Is the reproducibility of evidence in favour of homoeopathy proof of its activity or proof of the clinical trial's capacity to produce false-positive results?


Mutant cadherin affects epithelial morphogenesis and invasion, but not transformation.
MDCK cells were engineered to reversibly express mutant E-cadherin protein with a large extracellular deletion. Mutant cadherin overexpression reduced the expression of endogenous E- and K-cadherins in MDCK cells to negligible levels, resulting in decreased cell adhesion. Despite severe impairment of the cadherin adhesion system, cells overexpressing mutant E-cadherin formed fluid-filled cysts in collagen gel cultures and responded to hepatocyte growth factor/scatter factor (HGF/SF) that induced cellular extension formation with a frequency similar to that of control cysts. However, cells were shed from cyst walls into the lumen and into the collagen matrix prior to and during HGF/SF induced tubule extension. Despite the propensity for cell dissociation, MDCK cells lacking cadherin adhesion molecules were not capable of anchorage-independent growth in soft agar and cell proliferation rate was not affected. Thus, cadherin loss does not induce transformation, despite inducing an invasive phenotype, a later stage of tumor progression. These experiments are especially relevant to tumor progression in cells with altered E-cadherin expression, particularly tumor samples with identified E-cadherin extracellular domain genomic mutations.


Rate variation among nuclear genes and the age of polyploidy in Gossypium.
Molecular evolutionary rate variation in Gossypium (cotton) was characterized using sequence data for 48 nuclear genes from both genomes of allotetraploid cotton, models of its diploid progenitors, and an outgroup. Substitution rates varied widely among the 48 genes, with silent and replacement substitution levels varying from 0.018 to 0.162 and from 0.000 to 0.073, respectively, in comparisons between orthologous Gossypium and outgroup sequences. However, about 90% of the genes had silent substitution rates spanning a more narrow threefold range. Because there was no evidence of rate heterogeneity among lineages for any gene and because rates were highly correlated in independent tests, evolutionary rate is inferred to be a property of each gene or its genetic milieu rather than the clade to which it belongs. Evidence from approximately 200,000 nucleotides (40,000 per genome) suggests that polyploidy in Gossypium led to a modest enhancement in rates of nucleotide substitution. Phylogenetic analysis for each gene yielded the topology expected from organismal history, indicating an absence of gene conversion or recombination among homoeologs subsequent to allopolyploid formation. Using the mean synonymous substitution rate calculated across the 48 genes, allopolyploid cotton is estimated to have formed circa 1.5 million years ago (MYA), after divergence of the diploid progenitors about 6.7 MYA.


Timing of neonatal stoma closure: a survey of health professional perspectives and current practice.
Optimal timing for neonatal stoma closure remains unclear. In this study, we aimed to establish current practice and illustrate multidisciplinary perspectives on timing of stoma closure using an online survey sent to all 27 UK neonatal surgical units, as part of a research programme to determine the feasibility of a clinical trial comparing 'early' and 'late' stoma closure. 166 responses from all 27 units demonstrated concordance of opinion in target time for closure (6 weeks most commonly stated across scenarios), although there was a high variability in practice. A sizeable proportion (41%) of respondents use weight, rather than time, to determine when to close a neonatal stoma. Thematic analysis of free text responses identified nine key themes influencing decision-making; most related to nutrition, growth and stoma complications. These data provide an overview of current practice that is critical to informing an acceptable trial design.


Propofol versus midazolam for upper gastrointestinal endoscopy in cirrhotic patients: a meta-analysis of randomized controlled trials.
Sedation during gastrointestinal endoscopy is often achieved using propofol or midazolam in general population. However, impaired protein synthesis, altered drug metabolism, and compromised hepatic blood flow in patients with liver cirrhosis might affect the pharmacokinetics of sedatives, placing cirrhotic patients undergoing endoscopy at a greater risk of adverse events. The objective of this study was to assess comparative efficacies and safety of propofol and midazolam in cirrhotic patients undergoing endoscopy. Randomized, controlled trials comparing propofol with midazolam in cirrhotic patients undergoing gastrointestinal endoscopy were selected. We performed the meta-analysis, using a random-effect model, the Review Manager, Version 5.2, statistical software package (Cochrane Collaboration, Oxford, UK) according to the PRISMA guidelines. Five studies between 2003 and 2012, including 433 patients, were included. Propofol provided a shorter time to sedation (weight mean difference: -2.76 min, 95% confidence interval: -3.00 to -2.51) and a shorter recovery time (weight mean difference -6.17 min, 95% confidence interval: -6.81 to -5.54) than midazolam did. No intergroup difference in the incidence of hypotension, bradycardia, or hypoxemia was observed. Midazolam was associated with the deterioration of psychometric scores for a longer period than propofol. This meta-analysis suggests that Propofol sedation for endoscopy provides more rapid sedation and recovery than midazolam does. The risk of sedation-related side effects for propofol does not differ significantly from that of midazolam. The efficacy of propofol in cirrhotic patients undergoing endoscopy is superior to those of midazolam.


Increased osteoclast activity is associated with aggressiveness of osteosarcoma.
Osteosarcoma (OS) is a highly malignant primary skeletal tumor with a striking tendency to rapidly destroy the surrounding bone and metastasize, since metastases are frequently present at clinical onset. The basis for the aggressiveness of this tumor is largely unknown. However, recent studies in in vivo models indicate that the anti-osteolytic drugs, bisphosphonates, can inhibit the tumor local expansion and the formation of metastases. We further investigated the association between the presence of active osteoclasts and the aggressiveness of OS. We evaluated the presence of osteoclasts and the mRNA of different osteoclast-related genes in tumor biopsies from 16 OS patients and in three OS cell lines and the serum levels of bone resorption markers in the same series and in 28 other patients. Tumor-associated osteoclasts were found in 63 and 75% of cases by histological and mRNA analysis. Among different serum markers, only MMP-9 was significantly higher in OS cases (p=0.0001), whereas TRACP 5b was significantly higher in metastatic patients compared to nonmetastatic patients (p=0.0509). Serum TRACP 5b was significantly correlated to serum NTX (p<0.0001) and cathepsin K mRNA in tumor tissues (p=0.0153). In 8 patients we also analyzed TRACP 5b serum level at follow-up and we verified a significant decrease of TRACP 5b after primary tumor removal (p=0.0117). In conclusion, tumor-infiltrating osteoclasts are frequently found in OS and increased serum TRACP 5b levels and the presence of active osteoclast at primary sites were positively associated with tumor aggressiveness.


Development of 11-Plex MOL-PCR Assay for the Rapid Screening of Samples for Shiga Toxin-Producing Escherichia coli.
Strains of Shiga toxin-producing Escherichia coli (STEC) are a serious threat to the health, with approximately half of the STEC related food-borne illnesses attributable to contaminated beef. We developed an assay that was able to screen samples for several important STEC associated serogroups (O26, O45, O103, O104, O111, O121, O145, O157) and three major virulence factors (eae, stx 1 , stx 2) in a rapid and multiplexed format using the Multiplex oligonucleotide ligation-PCR (MOL-PCR) assay chemistry. This assay detected unique STEC DNA signatures and is meant to be used on samples from various sources related to beef production, providing a multiplex and high-throughput complement to the multiplex PCR assays currently in use. Multiplex oligonucleotide ligation-PCR (MOL-PCR) is a nucleic acid-based assay chemistry that relies on flow cytometry/image cytometry and multiplex microsphere arrays for the detection of nucleic acid-based signatures present in target agents. The STEC MOL-PCR assay provided greater than 90% analytical specificity across all sequence markers designed when tested against panels of DNA samples that represent different STEC serogroups and toxin gene profiles. This paper describes the development of the 11-plex assay and the results of its validation. This highly multiplexed, but more importantly dynamic and adaptable screening assay allows inclusion of additional signatures as they are identified in relation to public health. As the impact of STEC associated illness on public health is explored additional information on classification will be needed on single samples; thus, this assay can serve as the backbone for a complex screening system.


Signalling by insulin and IGF receptors: supporting acts and new players.
The signalling pathways utilised by insulin receptor (IR) and IGF receptor to transduce their diverse effects on cellular metabolism, growth and survival are well established in broad outline, but many details remain to be elucidated. Tyrosine phosphorylation of IR substrates and Shc initiates signalling via canonical phosphoinositide 3-kinase/Akt and Ras/MAP kinase pathways, which together mediate many of the actions of insulin and IGFs. However, a variety of additional substrates and scaffolds have been described that may play roles in modulating the canonical pathways or in specific biological responses. This review will focus on recent studies that have extended our understanding of insulin/IGF signalling pathways, and the elements that may contribute to specificity.


HSV-1 ICP0: An E3 Ubiquitin Ligase That Counteracts Host Intrinsic and Innate Immunity.
The herpes simplex virus type 1 (HSV-1) encoded E3 ubiquitin ligase, infected cell protein 0 (ICP0), is required for efficient lytic viral replication and regulates the switch between the lytic and latent states of HSV-1. As an E3 ubiquitin ligase, ICP0 directs the proteasomal degradation of several cellular targets, allowing the virus to counteract different cellular intrinsic and innate immune responses. In this review, we will focus on how ICP0's E3 ubiquitin ligase activity inactivates the host intrinsic defenses, such as nuclear domain 10 (ND10), SUMO, and the DNA damage response to HSV-1 infection. In addition, we will examine ICP0's capacity to impair the activation of interferon (innate) regulatory mediators that include IFI16 (IFN γ-inducible protein 16), MyD88 (myeloid differentiation factor 88), and Mal (MyD88 adaptor-like protein). We will also consider how ICP0 allows HSV-1 to evade activation of the NF-κB (nuclear factor kappa B) inflammatory signaling pathway. Finally, ICP0's paradoxical relationship with USP7 (ubiquitin specific protease 7) and its roles in intrinsic and innate immune responses to HSV-1 infection will be discussed.


Aspects of vitamin A.
Musgrave Park Hospital in 1942 was the site of an Anglo-American Vitamin A caper. A threatened court-martial was pre-empted. Subsequently the Queen's lecturer in Anatomy, JW Millen, who was the other lecturer to the first editor of this journal, RH Hunter, did much distinguished work. The neurological effects of Vitamin A were elucidated. Further work on cerebrospinal fluid (CSF), placenta, thalidomide and poliomyelitis led to the pre-eminence in applied anatomy and teratology of now Reader James Wilson Millen and Professors JD Boyd and WJ Hamilton, all Queen's Medical School graduates. Training of RH Hunter, JH Biggart and JD Boyd at Johns Hopkins University profoundly influenced these seminal discoveries. The Garretts, a family of Lisburn, County Down origin, saved Johns Hopkins Hospital and Medical School from financial disaster. The Garretts founded a commercial and mercantile empire that took control of the Baltimore and Ohio (B and O) Railroad and enabled the Garretts to dictate that women should be admitted to the Hopkins Medical School and Hospital on exactly the same terms as men. All women and men should already be university honours graduates. Winston S Churchill on his progress up and down the B and O main line in March 1946, recounted to President Harry S Truman and Harry Hopkins his mother's tales of the Garrett boys' adventures.


Controlled trial of natamycin in the treatment of allergic bronchopulmonary aspergillosis.
Allergic bronchopulmonary aspergillosis often requires treatment with oral corticosteroids to control the host response to Aspergillus fumigatus. In a double blind study 25 patients with allergic bronchopulmonary aspergillosis taking maintenance oral corticosteroids were randomly allocated to receive 5 mg natamycin or placebo by nebuliser twice daily for one year. The primary aim of the study was to assess the steroid sparing potential of natamycin. Standardised reductions in corticosteroid dosage were therefore undertaken every five weeks, unless clinically contraindicated. Five patients were withdrawn in the first four months: two (1 natamycin, 1 placebo) died, two (1 natamycin, 1 placebo) had suspected drug reactions, and one (natamycin) was non-compliant. The pretreatment characteristics of the 20 patients (10 in each group) who completed the study were similar, 17 (9 natamycin, 8 placebo) having evidence of recent disease activity. At the end of the study prednisolone dose had been reduced by a similar amount in each group (median natamycin 2.25 mg, placebo 2.5 mg). Evidence of disease activity during the study year (transient shadowing on the chest radiograph, blood eosinophilia, or increases in antibodies to A fumigatus, or any combination of these) was observed in similar numbers of patients in each group (5 natamycin, 7 placebo). There was no evidence that natamycin conferred benefit on these patients with allergic bronchopulmonary aspergillosis.


EURO-MUSCULUS/USPRM. Basic Scanning Protocols for Ankle and foot.
In this protocol, the patient/probe positionings, anatomical drawings and ultrasound images of commonly scanned ankle/foot structures are described. This practical guide is prepared (with an international consensus of several expert physiatrists) to serve as a uniform/standard approach especially for beginner sonographers.


Laparoscopic liver resection for hepatocellular carcinoma: a three-year study of 57 Patients.
Laparoscopic liver resection has become an increasingly popular operation but is still in relatively limited use. Here we evaluate the intermediate-term results of laparoscopic liver resection. Fifty-seven patients with HCC underwent laparoscopic liver resection at the Asan Medical Center. Data for all resections were recorded and analyzed retrospectively. Patient gender, age, preoperative laboratory data, presence of cirrhosis, blood loss, hospital stay length, pathology report, tumor site and size, resection type, resection margin, morbidity and mortality were assessed. The mean patient age ranged from 35-74 years and the mean tumor size from 0.8-5.5 cm. Tumors were located in the left lateral lobe, left medial lobe and right lobe. Left lateral sectionectomy was performed in 32 cases, partial hepatectomy in 11, left hepatectomy in 6, right hepatectomy in 2, laparoscopy-assisted right hepatectomy in 2 and laparoscopy-assisted right posterior sectionectomy in 4. Median operation time ranged from 95-380 min while median blood loss ranged from 150-800 mL. The mean resection margin was 2.08±1.68 cm with no in-hospital mortalities. Return to normal diet was achieved on average at 1.83±0.8 days; mean hospital stay ranged from 3 to 17 days. The 3-year overall and disease-free survival rates were 81% and 71%, respectively. Laparoscopic liver resection for HCC is feasible, safe and promising for a select group of patients. Its benefits include short hospital stays, rapid return to normal diet, full mobility and minimal morbidity with acceptable oncological parameters.


Maternal age, ethnicity and gestational diabetes mellitus.
in Australia, and globally, rates for gestational diabetes mellitus (GDM) have risen dramatically in recent decades. This is of concern as GDM is associated with adverse pregnancy outcomes and additional health-care costs. Factors linked to increasing incidence include older maternal age and non-Caucasian ethnicity. However, as yet, there is no clear consensus on the magnitude of effect associated with these factors in combination. This study therefore investigated the effect of maternal age and country/region of birth on GDM incidence. all women who gave birth in Victoria, Australia in 2005 and 2006 (n=133,359) were included in this population-based cross-sectional study. Stratified cross-tabulations were conducted to examine the incidence of GDM by maternal age group and country/region of birth. Primiparous women were further analysed separately from parous women. The proportion of women with GDM was reported, along with the χ(2) for linear trend. whilst women born outside Australia constituted just 24.6% of women giving birth during the study period, they accounted for 41.4% of GDM cases. The highest GDM incidence was seen among Asian women at 11.5%, compared with Australian born women at 3.7%. There was strong evidence that women born in all regions except North America were increasingly likely to develop GDM in pregnancies at older ages (p<0.001).On examining age related GDM trends by maternal region of birth, higher rates were seen across all regions studied but were most marked among women born in Asia and the Middle East. older maternal age and non-Australian birth increased a woman's risk of developing GDM and this increase was most evident among Asian women. As GDM is associated with adverse maternal and infant outcomes, it is important to explore ways of preventing GDM, and to put in place strategies to effectively manage GDM during pregnancy and to reduce the later risk of developing type 2 diabetes. Pregnancy presents midwives with a unique opportunity to provide education and to encourage dietary and behavioural modifications as women have repeated contact with the health system during this time.


Study protocol: evaluation of an online, father-inclusive, universal parenting intervention to reduce child externalising behaviours and improve parenting practices.
Parenting interventions that focus on enhancing the quality and consistency of parenting are effective for preventing and reducing externalising problems in children. There has been a recent shift towards online delivery of parenting interventions in order to increase their reach and impact on the population prevalence of child externalising problems. Parenting interventions have low rates of father participation yet research suggests that father involvement may be critical to the success of the intervention. Despite this, no online parenting interventions have been specifically developed to meet the needs and preferences of fathers, as well as mothers. This paper describes the protocol of a study examining the effectiveness of an online, father-inclusive parenting intervention called 'ParentWorks', which will be delivered as a universal intervention to Australian families. A single group clinical trial will be conducted to examine the effectiveness of ParentWorks for reducing child externalising problems and improving parenting, as well as to explore the impact of father engagement (in two-parent families) on child outcomes. Australian parents/caregivers with a child aged 2-16 years will be recruited. Participants will provide informed consent, complete pre-intervention measures and will then complete the intervention, which consists of five compulsory video modules and three optional modules. The primary outcomes for this study are changes in child externalising behaviour, positive and dysfunctional parenting practices and parental conflict, and the secondary outcome is changes in parental mental health. Demographic information, satisfaction with the intervention, and measures of parental engagement will also be collected. Questionnaire data will be collected at pre-intervention, post-intervention and three-month follow-up, as well as throughout the program. This paper describes the study protocol of a single group clinical trial of a national, online, father-inclusive parenting intervention. The results from this study could be used to inform public policy about providing support to parents of children with behaviour problems, and enhancing the engagement of fathers in parenting interventions. ACTRN12616001223426 , registered 05/09/2016.


Ultrasensitive electrogenerated chemiluminescence biosensor for the determination of mercury ion incorporating G4 PAMAM dendrimer and Hg(II)-specific oligonucleotide.
A novel electrogenerated chemiluminescence (ECL) biosensor for highly sensitive and selective detection of mercury ion was developed on the basis of mercury-specific oligonucleotide (MSO) served as a molecular recognition element and the ruthenium(II) complex (Ru1) as an ECL emitting species. The biosensor was fabricated on a glassy carbon electrode coated with a thin layer of single wall carbon nanotubes, where the ECL probe, NH(2)-(CH(2))(6)-oligo(ethylene oxide)(6)-MSO↔Dend-Ru1, was covalently attached. The Dend-Ru1 pendant was prepared by covalent coupling Ru1 with the 4th generation polyamidoamine dendrimer (Dend), in which each dendrimer contained 35 Ru1 units so that a large amplification of ECL signal was obtained. Upon binding of Hg(2+) to thymine (T) bases of the MSO, the T-Hg-T structure was formed, and the MSO changed from its linear shape to a "hairpin" configuration. Consequently, the Dend-Ru1 approached the electrode surface resulting in the increase of anodic ECL signal in the presence of the ECL coreactant tri-n-propylamine. The reported biosensor showed a high reproducibility and possessed long-term storage stability (92.3% initial ECL recovery over 30 day's storage). An extremely low detection limit of 2.4 pM and a large dynamic range of 7.0 pM to 50 nM Hg(2+) were obtained. An apparent binding constant of 1.6 × 10(9)M(-1) between Hg(2+) and the MSO was estimated using an ECL based extended Langmuir isotherm approach involving multilayer adsorption. Determination of Hg(2+) contents in real water samples was conducted and the data were consistent with the results from cold vapor atomic fluorescence spectroscopy.


Many faces of SMCHD1.
The chromatin scaffolding protein SMCHD1 (structural maintenance of chromosomes flexible hinge domain containing 1) was previously shown to have diverse roles in X-chromosome inactivation, imprinting and double-strand break repair, and mutations in SMCHD1 contribute to a type of muscular dystrophy. Now, development of the nose and eyes is added to its list of functions.


Prevalence and Factors Associated with Parents' Non-Intention to Vaccinate Their Children and Adolescents against COVID-19 in Latin America and the Caribbean.
We aimed to estimate the prevalence and factors associated with parents' non-intention to vaccinate their children and adolescents against COVID-19 in Latin America and the Caribbean (LAC). We performed a secondary analysis using a database generated by the University of Maryland and Facebook (Facebook, Inc., Menlo Park, CA, USA). We included adult (18 and over) Facebook users residing in LAC who responded to the survey between 20 May 2021 and 14 July 2021. We included sociodemographic characteristics, comorbidities, mental health, economic and food insecurity, compliance with mitigation strategies against COVID-19, and practices related to vaccination against this disease. We estimated the crude (cPR) and adjusted (aPR) prevalence ratios with their respective 95%CI. We analyzed a sample of 227,740 adults from 20 LAC countries. The prevalence of parents' non-intention to vaccinate their children and adolescents against COVID-19 was 7.8% (n = 15,196). An age above 35 years old, educational level above college, compliance with physical distancing, use of masks, having economic insecurity, having had COVID-19, anxiety symptoms, depressive symptoms, having a chronic condition or two or more comorbidities, and being vaccinated were associated with a lower prevalence of non-intention to vaccinate children and adolescents against COVID-19. Living in a town, a village, or a rural area was associated with a higher prevalence of non-intention to vaccinate children and adolescents against COVID-19. Approximately nine out of ten parents in LAC intended to vaccinate their children and adolescents against COVID-19. Our results allow for understanding parents' intentions to vaccinate children and adolescents and help promote and develop education strategies for national vaccination plans against COVID-19.


Seroprevalence and correlates of herpes simplex virus type 2 infection among young adults in Arkhangelsk, Northwest Russia: a population-based cross-sectional study.
Herpes simplex virus type 2 (HSV-2) infection is the most common cause of genital ulcer disease (GUD) worldwide. Mother to child transmission causes high morbidity and mortality among infants. Russia is on the brink of a generalized HIV-epidemic, but Arkhangelsk is still a low-prevalence area. HSV-2 infection is associated with a three-fold increased risk of HIV-infection. The evidence on the seroprevalence of HSV-2 in Russia is limited. The aim of this study was to assess HSV-2 seroprevalence and correlates among young adults in the city of Arkhangelsk. 1243 adults aged 18-39 years participated in a cross-sectional population-based study, recruited by a public opinion agency applying a quota sampling method to achieve a data set with similar age- and sex-distribution as the population in Arkhangelsk. All participants completed a standardized, self-administrated questionnaire and were tested for HSV-2. Associations between HSV-2 seropositivity and selected sociodemographic and behavioral factors, and self-reported history of sexually transmitted infections (STIs) were studied by multivariable logistic regression. HSV-2 seroprevalence was 18.8 %: 12.2 % (95 % confidence interval, CI 9.7-15.2) among men and 24.0 % (95 % CI 20.1-27.3) among women. Among men, HSV-2 positivity was associated with being divorced/widowed (OR = 2.85, 95 % CI 1.06-7.70), cohabitation (OR = 2.45, 95 % CI 1.07-5.62), and a history of STIs (OR = 2.11, 95 % CI 1.14-3.91). In women, HSV-2 positivity was associated with high income (OR = 3.11, 95 % CI 1.45-6.71) and having a lifetime number of sexual partners between 2 and 5 (OR = 2.72, 95 % CI 1.14-6.51), whereas sexual debut at age 18 years or older was inversely associated with the outcome (OR = 0.47, 95 % CI 0.31-0.72). In both sexes, increasing age was the strongest correlate of HSV-2 seropositivity in multivariable analyses. The HSV-2 seroprevalence was twice as high in women than in men and increased with age in both sexes, and similar to that reported from high-prevalence countries in Europe and the USA. The high prevalence of HSV-2 among women in childbearing age reveals the potential for HSV-2 transmission from mothers to infants and increased risk of acquisition HIV-infection; it also contributes to the burden GUD among both sexes. This emphasizes the public health implications of the HSV-2 epidemic in an urban population in North-West Russia.


The maize genome contains a helitron insertion.
The maize mutation sh2-7527 was isolated in a conventional maize breeding program in the 1970s. Although the mutant contains foreign sequences within the gene, the mutation is not attributable to an interchromosomal exchange or to a chromosomal inversion. Hence, the mutation was caused by an insertion. Sequences at the two Sh2 borders have not been scrambled or mutated, suggesting that the insertion is not caused by a catastrophic reshuffling of the maize genome. The insertion is large, at least 12 kb, and is highly repetitive in maize. As judged by hybridization, sorghum contains only one or a few copies of the element, whereas no hybridization was seen to the Arabidopsis genome. The insertion acts from a distance to alter the splicing of the sh2 pre-mRNA. Three distinct intron-bearing maize genes were found in the insertion. Of most significance, the insertion bears striking similarity to the recently described DNA helicase-bearing transposable elements termed HELITRONS: Like Helitrons, the inserted sequence of sh2-7527 is large, lacks terminal repeats, does not duplicate host sequences, and was inserted between a host dinucleotide AT. Like Helitrons, the maize element contains 5' TC and 3' CTRR termini as well as two short palindromic sequences near the 3' terminus that potentially can form a 20-bp hairpin. Although the maize element lacks sequence information for a DNA helicase, it does contain four exons with similarity to a plant DEAD box RNA helicase. A second Helitron insertion was found in the maize genomic database. These data strongly suggest an active Helitron in the present-day maize genome.


Expression of 8-oxoguanine glycosylase in human fetal membranes.
The most common DNA lesion generated by oxidative stress (OS) is 7, 8-dihydro-8-oxoguanine (8-oxoG) whose excision repair is performed by 8-oxoguanine glycosylase (OGG1). We investigated OGG1 expression changes in fetal membranes from spontaneous preterm birth (PTB) and preterm premature rupture of the membranes (pPROM) and its changes in vitro in normal fetal membranes exposed to OS inducer water-soluble cigarette smoke extract (CSE). DNA damage was determined in amnion cells treated with CSE by comet and FLARE assays. OGG1 mRNA expression and localization in fetal membranes from clinical specimens and in normal term membranes exposed to CSE were examined by QRT-PCR and by immunohistochemistry. DNA strand and base damage was seen in amnion cells exposed to CSE. OGG1 expression was 2.5-fold higher in PTB samples compared with pPROM (P = 0.045). No significant difference was seen between term and pPROM or PTB and term. CSE treatment showed a nonsignificant decrease in OGG1. OGG1 was localized to both amnion and chorion with less intense staining in pPROM and CSE-treated membranes. Increased OS-induced DNA damage predominated by 8-oxoG is likely to persist in fetal cells due to reduced availability of base excision repair enzyme OGG1. This can likely lead to fetal cell senescence associated with some adverse pregnancy outcome.


Uptake of taurocholic acid and cholic acid in isolated hepatocytes from rainbow trout.
The uptake of the bile acids cholate (CHA) and taurocholate (TCHA) was studied in isolated hepatocytes from rainbow trout (Oncorhynchus mykiss). Both CHA and TCHA were taken up in a concentration- and temperature-dependent manner with optimum temperature at 15 degrees C and a strikingly efficient uptake even at low temperatures (0-5 degrees C). The total uptake was a combination of a saturable [Michaelis-Menten constant (Km) for CHA, 20 microM; Km for TCHA, 19 microM] and a nonsaturable component. The maximal uptake rate of the saturable component was 416 and 805 pmol.mg protein-1.min-1 for CHA and TCHA, respectively. The uptake of both bile acids was shown to be energy dependent, since it was inhibited by the metabolic inhibitors antimycin A, oligomycin and carbonyl cyanide m-chlorophenylhydrazone. The uptake was clearly Na+ independent, since isosmotic replacement of extracellular Na+ by Li+, choline, or K+ did not inhibit the uptake. Furthermore, it seemed to be independent of the presence of extracellular Cl-, since it was not inhibited by replacement of Cl- with sodium gluconate. On the whole, our results show that the hepatocellular uptake of bile acids in rainbow trout is mediated by a Na(+)-independent carrier system, with characteristics resembling the corresponding transport component in mammalian hepatocytes, but with high efficiency even at low temperatures.


Cellular and humoral autoreactivity in idiopathic pulmonary fibrosis.
Idiopathic pulmonary fibrosis (IPF) is a morbid, refractory lung disorder with an unknown pathogenesis. To investigate potential adaptive immune mechanisms in IPF, we compared phenotypes and effector functions of peripheral CD4 T cells, autoantibody production, and proliferative responses of pulmonary hilar lymph node CD4 T cells to autologous lung extracts from afflicted patients and normals. Our results show that greater proportions of peripheral CD4 T lymphocytes in IPF subjects expressed MHC class II and CD154 (CD40L), and they more frequently elaborated TGF-beta1, IL-10, and TNF-alpha. Abnormal CD4 T cell clonal expansions were found in all IPF patients, and 82% of these subjects also had IgG autoantibodies against cellular Ags. IPF lung extracts stimulated proliferations of autologous CD4 T cells, unlike preparations from normals or those with other lung diseases, and the IPF proliferative responses were enhanced by repeated cycles of stimulation. Thus, CD4 T cells from IPF patients have characteristics typical of cell-mediated pathologic responses, including augmented effector functions, provision of facultative help for autoantibody production, oligoclonal expansions, and proliferations driven by an Ag present in diseased tissues. Recognition that an autoreactive immune process is present in IPF can productively focus efforts toward identifying the responsible Ag, and implementing more effective therapies.


Silent reading of direct versus indirect speech activates voice-selective areas in the auditory cortex.
In human communication, direct speech (e.g., Mary said: "I'm hungry") is perceived to be more vivid than indirect speech (e.g., Mary said [that] she was hungry). However, for silent reading, the representational consequences of this distinction are still unclear. Although many of us share the intuition of an "inner voice," particularly during silent reading of direct speech statements in text, there has been little direct empirical confirmation of this experience so far. Combining fMRI with eye tracking in human volunteers, we show that silent reading of direct versus indirect speech engenders differential brain activation in voice-selective areas of the auditory cortex. This suggests that readers are indeed more likely to engage in perceptual simulations (or spontaneous imagery) of the reported speaker's voice when reading direct speech as opposed to meaning-equivalent indirect speech statements as part of a more vivid representation of the former. Our results may be interpreted in line with embodied cognition and form a starting point for more sophisticated interdisciplinary research on the nature of auditory mental simulation during reading.


Bat Coronaviruses in China.
During the past two decades, three zoonotic coronaviruses have been identified as the cause of large-scale disease outbreaks⁻Severe Acute Respiratory Syndrome (SARS), Middle East Respiratory Syndrome (MERS), and Swine Acute Diarrhea Syndrome (SADS). SARS and MERS emerged in 2003 and 2012, respectively, and caused a worldwide pandemic that claimed thousands of human lives, while SADS struck the swine industry in 2017. They have common characteristics, such as they are all highly pathogenic to humans or livestock, their agents originated from bats, and two of them originated in China. Thus, it is highly likely that future SARS- or MERS-like coronavirus outbreaks will originate from bats, and there is an increased probability that this will occur in China. Therefore, the investigation of bat coronaviruses becomes an urgent issue for the detection of early warning signs, which in turn minimizes the impact of such future outbreaks in China. The purpose of the review is to summarize the current knowledge on viral diversity, reservoir hosts, and the geographical distributions of bat coronaviruses in China, and eventually we aim to predict virus hotspots and their cross-species transmission potential.


LYVE-1, a new homologue of the CD44 glycoprotein, is a lymph-specific receptor for hyaluronan.
The extracellular matrix glycosaminoglycan hyaluronan (HA) is an abundant component of skin and mesenchymal tissues where it facilitates cell migration during wound healing, inflammation, and embryonic morphogenesis. Both during normal tissue homeostasis and particularly after tissue injury, HA is mobilized from these sites through lymphatic vessels to the lymph nodes where it is degraded before entering the circulation for rapid uptake by the liver. Currently, however, the identities of HA binding molecules which control this pathway are unknown. Here we describe the first such molecule, LYVE-1, which we have identified as a major receptor for HA on the lymph vessel wall. The deduced amino acid sequence of LYVE-1 predicts a 322-residue type I integral membrane polypeptide 41% similar to the CD44 HA receptor with a 212-residue extracellular domain containing a single Link module the prototypic HA binding domain of the Link protein superfamily. Like CD44, the LYVE-1 molecule binds both soluble and immobilized HA. However, unlike CD44, the LYVE-1 molecule colocalizes with HA on the luminal face of the lymph vessel wall and is completely absent from blood vessels. Hence, LYVE-1 is the first lymph-specific HA receptor to be characterized and is a uniquely powerful marker for lymph vessels themselves.


Diabetes Mellitus is Associated With Mortality in Acute Pancreatitis.
Predicting severe pancreatitis is important for early aggressive management of patients with acute pancreatitis (AP). Despite the established role of diabetes mellitus (DM) in the risk of AP, the impact of DM on the clinical outcome in AP has not been fully elucidated. The objective of this study was to assess the risk of mortality and severity in AP among patients with type-2 DM. Patients diagnosed with first attacks of AP were enrolled from January 2013 to June 2015. A total of 201 patients (63.2% male, mean age, 59.4 y) with AP were included. Etiologies included gallstones (51.2%), alcohol (37.3%), hypertriglyceridemia (2%), and idiopathic causes (9.5%). There were 54 AP patients (26.9%) with type-2 DM. Severity indices in AP, such as Atlanta Classification (severe), Ranson score, and Bedside Index of Severity in Acute Pancreatitis, were higher in subjects with DM than those without DM. Prevalence of intensive care unit admission and mortality were higher in AP patients with DM compared with those without DM. The association between DM and increased risk of mortality in AP remained statistically significant even after adjustments for confounding factors and Atlanta Classification (odds ratio, 7.76, 95% confidence interval, 1.26-47.63, P=0.027). Type-2 DM was associated with severity and increased mortality in patients with AP. Our findings provide evidence of the potential role of DM in the pathogenesis and management of severe AP.


PsychDT Working Group: Report Psychosocial Aspects of Artificial Pancreas Systems.
Diabetes technology is a cornerstone of diabetes management in the 21st century, with advances in available devices over recent years playing a central role in the way that health care has progressed. Psychosocial interventions have been shown to have a positive impact on glycemic control, reduce psychological distress and reduce costs of health care. Addressing and improving psychosocial outcomes that complement biomedical improvements and looking to the future are crucial to enhance patient acceptance of artificial pancreas (AP) systems. To achieve closer collaboration and comparability across different AP research trials, a working group was established. Existing measures fail to adequately capture the extent to which human and psychological factors play a role in the uptake and efficient use of AP systems. Understanding these factors will ultimately lead to the most benefit for users. Reliable measures of the psychosocial impact of AP systems for users is crucial to ensure that (1) regulatory authorities are able to robustly consider these aspects as part of their approval process, (2) government and private payers are able to factor these aspects into their decisions regarding reimbursement, and (3) persons with diabetes maximize benefits in terms of both glycemic control and quality of life to minimize the burden of diabetes in everyday life. This working group will serve as a platform to foster exchange, identify research needs, and guide and initiate collaborative research laying the groundwork for optimal utilization of diabetes technology in clinical diabetes care. A close collaboration among all key stakeholders is crucial to ensure that devices are designed, trialed, approved, and provided with minimal user burden and maximum beneficial effect.


The adaptor molecule TIRAP provides signalling specificity for Toll-like receptors.
Mammalian Toll-like receptors (TLRs) function as sensors of infection and induce the activation of innate and adaptive immune responses. Upon recognizing conserved pathogen-associated molecular products, TLRs activate host defence responses through their intracellular signalling domain, the Toll/interleukin-1 receptor (TIR) domain, and the downstream adaptor protein MyD88 (refs 1-3). Although members of the TLR and the interleukin-1 (IL-1) receptor families all signal through MyD88, the signalling pathways induced by individual receptors differ. TIRAP, an adaptor protein in the TLR signalling pathway, has been identified and shown to function downstream of TLR4 (refs 4, 5). Here we report the generation of mice deficient in the Tirap gene. TIRAP-deficient mice respond normally to the TLR5, TLR7 and TLR9 ligands, as well as to IL-1 and IL-18, but have defects in cytokine production and in activation of the nuclear factor NF-kappaB and mitogen-activated protein kinases in response to lipopolysaccharide, a ligand for TLR4. In addition, TIRAP-deficient mice are also impaired in their responses to ligands for TLR2, TLR1 and TLR6. Thus, TIRAP is differentially involved in signalling by members of the TLR family and may account for specificity in the downstream signalling of individual TLRs.


Exposure rates of wrapped and unwrapped orbital implants following enucleation.
To compare the complication rate of porous polyethylene orbital motility implants with solid acrylic implants following enucleation and identify possible risk factors. The authors retrospectively reviewed the charts of a total of 117 consecutive enucleations performed at the University of Illinois at Chicago between March 28, 1994, and May 28, 1999. Data obtained included patient demographics, surgical indication, implant type, attending surgeon, surgical technique, and any reported complications. The primary outcome was presence or absence of implant exposure at the final recorded visit. Of the 117 identified cases, 29 were eliminated due to insufficient follow-up data. Of the 88 remaining cases, 48 patients received porous implants and 40 received solid acrylic implants. Implant exposure developed in four cases. All exposures occurred in unwrapped porous polyethylene implants (n=2) or porous polyethylene implants wrapped in absorbable material (n=2). All exposures occurred in patients younger than 18 years of age, and 75% occurred early after trauma-associated enucleation surgery. The exposure rate of porous polyethylene implants in this study (9%) was found to be comparable to published rates for hydroxyapatite implants. There were no exposures of unwrapped solid acrylic spheres. Unwrapped porous implants in pediatric patients or following trauma-related enucleation may represent an increased risk for postoperative implant exposure. Absorbable wrapping of porous implants may carry the same risk for exposure as no wrapping. Porous implants wrapped in durable material appear to be as safe as solid acrylic spheres.


Imaging in patients with suspected acute heart failure: timeline approach position statement on behalf of the Heart Failure Association of the European Society of Cardiology.
Acute heart failure is one of the main diagnostic and therapeutic challenges in clinical practice due to a non-specific clinical manifestation and the urgent need for timely and tailored management at the same time. In this position statement, the Heart Failure Association aims to systematize the use of various imaging methods in accordance with the timeline of acute heart failure care proposed in the recent guidelines of the European Society of Cardiology. During the first hours of admission the point-of-care focused cardiac and lung ultrasound examination is an invaluable tool for rapid differential diagnosis of acute dyspnoea, which is highly feasible and relatively easy to learn. Several portable and stationary imaging modalities are being increasingly used for the evaluation of cardiac structure and function, haemodynamic and volume status, precipitating myocardial ischaemia or valvular abnormalities, and systemic and pulmonary congestion. This paper emphasizes the central role of the full echocardiographic examination in the identification of heart failure aetiology, severity of cardiac dysfunction, indications for specific heart failure therapy, and risk stratification. Correct evaluation of cardiac filling pressures and accurate prognostication may help to prevent unscheduled short-term readmission. Alternative advanced imaging modalities should be considered to assist patient management in the pre- and post-discharge phase, including cardiac magnetic resonance, computed tomography, nuclear studies, and coronary angiography. The Heart Failure Association addresses this paper to the wide spectrum of acute care and heart failure specialists, highlighting the value of all available imaging techniques at specific stages and in common clinical scenarios of acute heart failure.


The ten-year survival of the Birmingham hip resurfacing: an independent series.
Recent events have highlighted the importance of implant design for survival and wear-related complications following metal-on-metal hip resurfacing arthroplasty. The mid-term survival of the most widely used implant, the Birmingham Hip Resurfacing (BHR), has been described by its designers. The aim of this study was to report the ten-year survival and patient-reported functional outcome of the BHR from an independent centre. In this cohort of 554 patients (646 BHRs) with a mean age of 51.9 years (16.5 to 81.5) followed for a mean of eight years (1 to 12), the survival and patient-reported functional outcome depended on gender and the size of the implant. In female hips (n = 267) the ten-year survival was 74% (95% confidence interval (CI) 83 to 91), the ten-year revision rate for pseudotumour was 7%, the mean Oxford hip score (OHS) was 43 (SD 8) and the mean UCLA activity score was 6.4 (SD 2). In male hips (n = 379) the ten-year survival was 95% (95% CI 92.0 to 97.4), the ten-year revision rate for pseudotumour was 1.7%, the mean OHS was 45 (SD 6) and the mean UCLA score was 7.6 (SD 2). In the most demanding subgroup, comprising male patients aged < 50 years treated for primary osteoarthritis, the survival was 99% (95% CI 97 to 100). This study supports the ongoing use of resurfacing in young active men, who are a subgroup of patients who tend to have problems with conventional THR. In contrast, the results in women have been poor and we do not recommend metal-on-metal resurfacing in women. Continuous follow-up is recommended because of the increasing incidence of pseudotumour with the passage of time.


Mutation of the catalytic domain of the foamy virus reverse transcriptase leads to loss of processivity and infectivity.
Foamy virus (FV) replication is resistant to most nucleoside analog reverse transcriptase (RT) inhibitors. In an attempt to create a 2',3'-dideoxy-3'-thiacytidine (3TC)-sensitive virus, the second residue in the highly conserved YXDD motif of simian foamy virus-chimpanzee (human isolate) [SFVcpz(hu)] RT was changed from Val (V) to Met (M). Unexpectedly, the resultant virus, SFVcpz(hu) RT-V313M, replicated poorly, and Met rapidly reverted to Val. Despite the presence of approximately 50% of wild-type RT activity in RT-V313M virions, full-length DNA products were not detected in transfected cells. Using purified recombinant enzymes, we found that the wild-type FV RT is significantly more processive than human immunodeficiency virus type 1 RT. However, the V313M mutant has about 40% of the wild-type level of FV RT activity and has a lower processivity than the wild-type FV enzyme. The V313M mutant RT is also relatively resistant to 3TC. These results suggest that the decrease in RT activity and processivity of FV RT-V313M prevents completion of reverse transcription and greatly diminishes viral replication.


Mice deficient of Lats1 develop soft-tissue sarcomas, ovarian tumours and pituitary dysfunction.
The lats gene has been identified as a tumour suppressor in Drosophila melanogaster using mosaic screens. Mosaic flies carrying somatic cells that are mutant for lats develop large tumours in many organs. The human LATS1 homologue rescues embryonic lethality and inhibits tumour growth in lats mutant flies, demonstrating the functional conservation of this gene. Biochemical and genetic analyses have revealed that LATS1 functions as a negative regulator of CDC2 (ref. 3). These data suggest that mammalian LATS1 may have a role in tumorigenesis. To elucidate the function of mammalian LATS1, we have generated Lats1-/- mice. Lats1-/- animals exhibit a lack of mammary gland development, infertility and growth retardation. Accompanying these defects are hyperplastic changes in the pituitary and decreased serum hormone levels. The reproductive hormone defects of Lats1-/- mice are reminiscent of isolated LH-hypogonadotropic hypogonadism and corpus luteum insufficiency in humans. Furthermore, Lats1-/- mice develop soft-tissue sarcomas and ovarian stromal cell tumours and are highly sensitive to carcinogenic treatments. Our data demonstrate a role for Lats1 in mammalian tumorigenesis and specific endocrine dysfunction.


Combined familial C7 and C4B deficiency in an adult with meningococcal disease.
A case of meningococcal septicaemia is reported in an adult with a deficiency of the seventh component of complement combined with a deficiency of the B locus product of C4. A family study demonstrated that the two deficiencies were not linked. This is the first time that the individual alleles of C4 were determined in a patient with a deficiency of one component of the terminal pathway. It is possible that the heterogeneous clinical picture of a terminal pathway deficiency may, in part, be explained by the co-existence of other subtle complement defects.


Six-Sigma Quality Management of Additive Manufacturing.
Quality is a key determinant in deploying new processes, products, or services and influences the adoption of emerging manufacturing technologies. The advent of additive manufacturing (AM) as a manufacturing process has the potential to revolutionize a host of enterprise-related functions from production to the supply chain. The unprecedented level of design flexibility and expanded functionality offered by AM, coupled with greatly reduced lead times, can potentially pave the way for mass customization. However, widespread application of AM is currently hampered by technical challenges in process repeatability and quality management. The breakthrough effect of six sigma (6S) has been demonstrated in traditional manufacturing industries (e.g., semiconductor and automotive industries) in the context of quality planning, control, and improvement through the intensive use of data, statistics, and optimization. 6S entails a data-driven DMAIC methodology of five steps-define, measure, analyze, improve, and control. Notwithstanding the sustained successes of the 6S knowledge body in a variety of established industries ranging from manufacturing, healthcare, logistics, and beyond, there is a dearth of concentrated application of 6S quality management approaches in the context of AM. In this article, we propose to design, develop, and implement the new DMAIC methodology for the 6S quality management of AM. First, we define the specific quality challenges arising from AM layerwise fabrication and mass customization (even one-of-a-kind production). Second, we present a review of AM metrology and sensing techniques, from materials through design, process, and environment, to postbuild inspection. Third, we contextualize a framework for realizing the full potential of data from AM systems and emphasize the need for analytical methods and tools. We propose and delineate the utility of new data-driven analytical methods, including deep learning, machine learning, and network science, to characterize and model the interrelationships between engineering design, machine setting, process variability, and final build quality. Fourth, we present the methodologies of ontology analytics, design of experiments (DOE), and simulation analysis for AM system improvements. In closing, new process control approaches are discussed to optimize the action plans, once an anomaly is detected, with specific consideration of lead time and energy consumption. We posit that this work will catalyze more in-depth investigations and multidisciplinary research efforts to accelerate the application of 6S quality management in AM.


Prevalence and trends of overweight among preschool children in developing countries.
Obesity during childhood is a matter of growing concern. Several reports show increasing rates of obesity in developed countries, whereas the extent of the problem in developing countries remains unknown. The aim of this study was to fill this gap by quantifying the prevalence and trends of overweight among preschool children in developing countries. One hundred sixty nationally representative cross-sectional surveys from 94 countries were analyzed in a standardized way to allow comparisons across countries and over time. Overweight was defined as a weight-for-height >2 SDs from the National Center for Health Statistics/World Health Organization international reference median. Prevalences of wasted children (< -2 SDs) are also presented to enable comparisons between both ends of the distribution. The global prevalence of overweight was 3.3%. Some countries and regions, however, had considerably higher rates, and overweight was shown to increase in 16 of 38 countries with trend data. Countries with the highest prevalences of overweight are located mainly in the Middle East, North Africa, and Latin America. Rates of wasting were generally higher than those of overweight; Africa and Asia had wasting rates 2.5-3.5 times higher than overweight rates. Countries with high wasting rates tended to have low overweight rates and vice versa. These estimates show that attention should be paid to monitoring levels and trends of overweight in children. This, however, should not be done at the expense of decreasing international commitments to alleviating undernutrition. The data presented confirm that undernutrition remains a major public health problem worldwide.


Population toxicokinetics of benzene.
In assessing the distribution and metabolism of toxic compounds in the body, measurements are not always feasible for ethical or technical reasons. Computer modeling offers a reasonable alternative, but the variability and complexity of biological systems pose unique challenges in model building and adjustment. Recent tools from population pharmacokinetics, Bayesian statistical inference, and physiological modeling can be brought together to solve these problems. As an example, we modeled the distribution and metabolism of benzene in humans. We derive statistical distributions for the parameters of a physiological model of benzene, on the basis of existing data. The model adequately fits both prior physiological information and experimental data. An estimate of the relationship between benzene exposure (up to 10 ppm) and fraction metabolized in the bone marrow is obtained and is shown to be linear for the subjects studied. Our median population estimate for the fraction of benzene metabolized, independent of exposure levels, is 52% (90% confidence interval, 47-67%). At levels approaching occupational inhalation exposure (continuous 1 ppm exposure), the estimated quantity metabolized in the bone marrow ranges from 2 to 40 mg/day.


The drug efficacy and adverse reactions in a mouse model of oral squamous cell carcinoma treated with oxaliplatin at different time points during a day.
Recent studies have shown that the growth and proliferation of cancer cells in vivo exhibit circadian rhythm, and the efficacy and adverse reactions of platinum-based anticancer drugs administered at different times of the day vary significantly on colon cancer. However, since the circadian rhythms of growth and proliferation of various cancer cells often differ, the question of whether the administration of platinum anticancer drugs at different times of the day exerts significantly different efficacy and adverse effects on oral cancers remains to be elucidated. This study has compared the efficacy and adverse effects of oxaliplatin (L-OHP) administration at different times during a day on oral squamous cell carcinoma in mice and has analyzed cellular circadian rhythms. The mouse model for oral squamous cell carcinoma was established in 75 nude mice, housed in a 12 hour light/12 hour dark cycle environment. The mice were randomly divided into five groups; four experimental groups were intravenously injected with L-OHP at four time points within a 24-hour period (4, 10, 16, and 22 hours after lights on [HALO]). The control group was intravenously injected with the same volume of saline. Treatment efficacy and adverse reactions were compared on the seventh day after the injection, at 22 HALO. The existence of circadian rhythms was determined by cosine analysis. Only injections of L-OHP at 16 and 22 HALO significantly prolonged animal survival time. The adverse reactions in mice injected with L-OHP at 16 and 22 HALO were significantly less than those observed in mice administered L-OHP at 4 and 10 HALO. The cosine fitting curve showed that the survival time and adverse reactions exhibited circadian rhythm. The time factor should be considered when treating patients with oral squamous cell carcinoma with L-OHP in order to achieve better efficacy, reduce the adverse reactions, and improve the patients' survival time and quality of life.


Rapid and cost-effective gene targeting in rat embryonic stem cells by TALENs.
The rat is the preferred animal model in many areas of biomedical research and drug development. Genetic manipulation in rats has lagged behind that in mice due to the lack of efficient gene targeting tools. Previously, we generated a knockout rat via conventional homologous recombination in rat embryonic stem (ES) cells. Here, we show that efficient gene targeting in rat ES cells can be achieved quickly through transcription activator-like effector nuclease (TALEN)-mediated DNA double-strand breaks. Using the Golden Gate cloning technique, we constructed a pair of TALEN targeting vectors for the gene of interest in 5 days. After gene transfection, the targeted rat ES cell colonies were isolated, screened, and confirmed by PCR without the need of drug selection. Our results suggest that TALEN-mediated gene targeting is a superior means of establishing genetically modified rat ES cell lines with high efficiency and short turnaround time.


The bigger, the better? Volume measurements of parasites and hosts: Parasitic barnacles (Cirripedia, Rhizocephala) and their decapod hosts.
Rhizocephala, a group of parasitic castrators of other crustaceans, shows remarkable morphological adaptations to their lifestyle. The adult female parasite consists of a body that can be differentiated into two distinct regions: a sac-like structure containing the reproductive organs (the externa), and a trophic, root like system situated inside the hosts body (the interna). Parasitism results in the castration of their hosts, achieved by absorbing the entire reproductive energy of the host. Thus, the ratio of the host and parasite sizes is crucial for the understanding of the parasite's energetic cost. Using advanced imaging methods (micro-CT in conjunction with 3D modeling), we measured the volume of parasitic structures (externa, interna, egg mass, egg number, visceral mass) and the volume of the entire host. Our results show positive correlations between the volume of (1) entire rhizocephalan (externa + interna) and host body, (2) rhizocephalan externa and host body, (3) rhizocephalan visceral mass and rhizocephalan body, (4) egg mass and rhizocephalan externa, (5) rhizocephalan egg mass and their egg number. Comparing the rhizocephalan Sylon hippolytes, a parasite of caridean shrimps, and representatives of Peltogaster, parasites of hermit crabs, we could match their different traits on a reconstructed relationship. With this study we add new and significant information to our global understanding of the evolution of parasitic castrators, of interactions between a parasitic castrator and its host and of different parasitic strategies within parasitic castrators exemplified by rhizocephalans.


A cross-sectional study of emergency care utilization and associated costs of violent-related (assault) injuries in the United States.
Violent-related (assault) injuries are a leading cause of death and disability in the United States. Many violent injury victims seek treatment in the emergency department (ED). Our objectives were to (1) estimate rates of violent-related injuries evaluated in United States EDs, (2) estimate linear trends in ED visits for violent-related injuries from 2000 to 2010, and (3) to determine the associated health care and work-loss costs. We examined adults 18 years and older from a nationally representative survey (the National Hospital Ambulatory Medical Care Survey) of ED visits, from 2000 to 2010. Violent injury was defined using International Classification of Diseases-9th Rev.-Clinical Modification, diagnosis and mechanism of injury codes. We calculated rates of ED visits for violent injuries. Medical and work-loss costs accrued by these injuries were calculated for 2005, inflation-adjusted to 2011 dollars using the WISQARS Cost of Injury Reports. An annual average of 1.4 million adults were treated for violent injuries in EDs from 2000 to 2010, comprising 1.6% (95% confidence interval, 1.5%-1.6%) of all US adult ED visits. Young adults (18-25 years), men, nonwhites, uninsured or publically insured patients, and those residing in high poverty urban areas were at increased risk for ED visits for violent injury. The 1-year, inflation-adjusted medical and work-loss cost of violent-inflicted injuries in adults in the United States was US $49.5 billion. Violent injuries account for over one million ED visits annually among adults, with no change in rates over the past decade. Young black men are at especially increased risk for ED visits for violent injuries. Overall, violent-related injuries resulted in substantial financial and societal costs. Epidemiological study, level III.


The misuse of BLUP in ecology and evolution.
Best linear unbiased prediction (BLUP) is a method for obtaining point estimates of a random effect in a mixed effect model. Over the past decade it has been used extensively in ecology and evolutionary biology to predict individual breeding values and reaction norms. These predictions have been used to infer natural selection, evolutionary change, spatial-genetic patterns, individual reaction norms, and frailties. In this article we show analytically and through simulation and example why BLUP often gives anticonservative and biased estimates of evolutionary and ecological parameters. Although some concerns with BLUP methodology have been voiced before, the scale and breadth of the problems have probably not been widely appreciated. Bias arises because BLUPs are often used to estimate effects that are not explicitly accounted for in the model used to make the predictions. In these cases, predicted breeding values will often say more about phenotypic patterns than the genetic patterns of interest. An additional problem is that BLUPs are point estimates of quantities that are usually known with little certainty. Failure to account for this uncertainty in subsequent tests can lead to both bias and extreme anticonservatism. We demonstrate that restricted maximum likelihood and Bayesian solutions exist for these problems and show how unbiased and powerful tests can be derived that adequately quantify uncertainty. Of particular utility is a new test for detecting evolutionary change that not only accounts for prediction error in breeding values but also accounts for drift. To illustrate the problem, we apply these tests to long-term data on the Soay sheep (Ovis aries) and the great tit (Parus major) and show that previously reported temporal trends in breeding values are not supported.