Acute appendicitis in patients with end-stage renal disease.
Acute appendicitis in patients with end-stage renal disease (ESRD) poses a diagnostic challenge. Delayed surgery can contribute to higher morbidity and mortality rates. However, few studies have evaluated this disease among ESRD patients. Our study focused on the lack of data on the incidence and risk factors of acute appendicitis among ESRD patients and compared the outcomes in patients who underwent different dialysis modalities. This national survey was conducted between 1997 and 2005 and included ESRD patients identified from the Taiwan National Health Insurance database. The incidence rate of acute appendicitis in ESRD patients was compared with that in randomly selected age-, sex-, and Charlson comorbidity score-matched non-dialysis controls. A Cox regression hazard model was used to identify risk factors. Among 59,781 incident ESRD patients, matched one-to-one with controls, there were 328 events of acute appendicitis. The incidence rate of 16.9 per 10,000 person-years in the ESRD cohort was higher than that in the control cohort (p = 0.003). The independent risk factors were atrial fibrillation (hazard ratio [HR], 2.08), severe liver disease (HR, 1.74), diabetes mellitus (HR, 1.58), and hemodialysis (HR, 1.74). Compared with the control cohort, subsequent perforation and mortality rates of acute appendicitis were also higher in the ESRD cohorts. There was no effect of dialysis modality on the patient outcomes. ESRD patients had a higher risk for acute appendicitis and poorer outcomes than non-dialysis populations. A careful examination of ESRD patients presenting with atypical abdominal pain to avoid misdiagnosis is extremely important to prevent delayed surgery.


A mineralogical study in contrasts: highly mineralized whale rostrum and human enamel.
The outermost enamel of the human tooth and the rostrum of the whale Mesoplodon densirostris are two highly mineralized tissues that contain over 95 wt.% mineral, i.e., bioapatite. However, the same mineral type (carbonated hydroxylapatite) does not yield the same material properties, as revealed by Raman spectroscopy, scanning electron microscopy, electron microprobe analysis, and synchrotron X-ray diffraction analysis. Overall, the outermost enamel of a tooth has more homogeneous physical and chemical features than the rostrum. Chemical comparison of rostrum and enamel shows bioapatite in the rostrum to be enriched in Na, Mg, CO3, and S, whereas the outermost enamel shows only a slightly enriched Cl concentration. Morphologically, mineral rods (at tens of μm scale), crystallites and prisms (at μm and sub-μm scale), and platelets (at tens of nm scale) all demonstrate less organized texture in the rostrum than in enamel. Such contrasts between two mineralized tissues suggest distinct pathways of biomineralization, e.g., the nature of the equilibrium between mineral and body fluid. This study illustrates the remarkable flexibility of the apatite mineral structure to match its chemical and physical properties to specific biological needs within the same animal or between species.


Electron microscopic evidence for a mucin-like region in chick muscle alpha-dystroglycan.
alpha-Dystroglycan has been isolated from chicken cardiac muscle and its molecular weight was estimated to be approximately 135 kDa. The avian protein interacts with murine Engelbreth-Holm-Swarm (EHS) tumor laminin via interaction with the C-terminal LG4 and LG5 domains (fragment E3) of the laminin alpha-chain. This laminin binding is calcium-dependent and can be competed by heparin. Electron microscopy investigation on the shape of alpha-dystroglycan suggests that the core protein consists of two roughly globular domains connected by a segment which most likely corresponds to a mucin-like central region also predicted by sequence analysis on mammalian isoforms. This segment may act as a spacer in the dystrophin-associated glycoproteins complex exposing the N-terminal domain of alpha-dystroglycan to laminin in the extracellular space.


In vivo single molecular imaging and sentinel node navigation by nanotechnology for molecular targeting drug-delivery systems and tailor-made medicine.
The recent advances in nanotechnology have a great potential to improve the prevention, diagnosis, and treatment of human diseases. Nanomaterials for medical applications are expected to grasp pharmacokinetics and the toxicity for application to medical treatment on the aspect of safety of the nanomaterials and nanodevices. We describe a generation of CdSe nanoparticles [quantum dots (QDs)] conjugated with monoclonal anti-HER2 antibody (Trastuzumab), for single molecular in vivo imaging of breast cancer cells. We established a high-resolution in vivo 3D microscopic system for a novel imaging method at the molecular level. The cancer cells expressing HER2 protein were visualized by the nanoparticles in vivo at subcellular resolution, suggesting future utilization of the system in medical applications to improve drug-delivery systems to target the primary and metastatic tumors for made-to-order treatment. We also describe sentinel node navigation using fluorescent nanoparticles for breast cancer surgery in experimental model, which have shown the potential to be an alternative to existing tracers in the detection of the sentinel node if we select the appropriate particle size and wavelength. Future innovation in cancer imaging by nanotechnology and novel measurement technology will provide great improvement, not only in the clinical field but also in basic medical science for the development of medicine.


Intrabodies based on intracellular capture frameworks that bind the RAS protein with high affinity and impair oncogenic transformation.
We have applied in vivo intracellular antibody capture (IAC) technology to isolate human intrabodies which bind to the oncogenic RAS protein. IAC facilitates the capture of antibody fragments, in this case single-chain Fvs (scFvs), which tolerate reducing environments, such as the cytoplasm of cancer cells. Three anti-RAS scFvs with different affinity, solubility and intracellular binding activity were characterized. The anti-RAS scFvs with highest affinity were expressed relatively poorly in mammalian cells, and greater soluble expression was achieved by mutating the antibody framework to canonical consensus scaffolds, previously derived from IAC, without losing antigen specificity. Mutagenesis experiments showed that the consensus scaffolds are functional as intrabody fragments without an intra-domain disulfide bond. Furthermore, we could convert an intrabody which does not bind RAS in mammalian cells into a high-affinity reagent capable of inhibiting RAS-mediated NIH 3T3 transformation by exchanging VH and VL complementarity-determining regions onto its consensus scaffold. These data show that the consensus scaffold is a robust framework by which to improve intrabody function.


Inhalable constituents of thirdhand tobacco smoke: chemical characterization and health impact considerations.
Tobacco smoke residues lingering in the indoor environment, also termed thirdhand smoke (THS), can be a source of long-term exposure to harmful pollutants. THS composition is affected by chemical transformations and by air-surface partitioning over time scales of minutes to months. This study identified and quantified airborne THS pollutants available for respiratory exposure, identified potential environmental tracers, and estimated health impacts to nonsmokers. In a ventilated 18 m(3) laboratory chamber, six cigarettes were machine-smoked, and levels of particulate matter (PM2.5) and 58 volatile organic compounds (VOCs) were monitored during an aging period of 18 h. Results were compared with field measurements taken in a smoker's home 8 h after the last cigarette had been smoked. Initial chamber levels of individual VOCs in freshly emitted secondhand smoke (SHS) were in the range of 1-300 μg m(-3). The commonly used SHS tracers 3-ethenylpyridine (3-EP) and nicotine were no longer present in the gas phase after 2 h, likely due mostly to sorption to surfaces. By contrast, other VOCs persisted in the gas phase for at least 18 h, particularly furans, carbonyls, and nitriles. The concentration ratio of acetonitrile to 3-EP increased substantially with aging. This ratio may provide a useful metric for differentiating freshly emitted (SHS) from aged smoke (THS). Among the 29 VOCs detected in the smoker's home at moderate to high concentrations, 18 compounds were also detected in simultaneously sampled outdoor air, but acetonitrile, 2-methyl furan, and 2,5-dimethyl furan appeared to be specific to cigarette smoke. The levels of acrolein, methacrolein, and acrylonitrile exceeded concentrations considered harmful by the State of California. An initial exposure and impact assessment was conducted for a subset of pollutants by computing disability-adjusted life years lost, using available toxicological and epidemiological information. Exposure to PM2.5 contributed to more than 90% of the predicted harm. Acrolein, furan, acrylonitrile, and 1,3-butadiene were considered to be the most harmful VOCs. Depending on which criteria are used to establish the separation between SHS and THS, 5-60% of the predicted health damage could be attributed to THS exposure. Benefits and limitations of this approach are discussed.


Receptor-mediated endocytosis of α-galactosidase A in human podocytes in Fabry disease.
Injury to the glomerular podocyte is a key mechanism in human glomerular disease and podocyte repair is an important therapeutic target. In Fabry disease, podocyte injury is caused by the intracellular accumulation of globotriaosylceramide. This study identifies in the human podocyte three endocytic receptors, mannose 6-phosphate/insulin-like growth II receptor, megalin, and sortilin and demonstrates their drug delivery capabilities for enzyme replacement therapy. Sortilin, a novel α-galactosidase A binding protein, reveals a predominant intracellular expression but also surface expression in the podocyte. The present study provides the rationale for the renal effect of treatment with α-galactosidase A and identifies potential pathways for future non-carbohydrate based drug delivery to the kidney podocyte and other potential affected organs.


Methicillin-sensitive and methicillin-resistant Staphylococcus aureus: preventing surgical site infections following plastic surgery.
The reader is presumed to have a broad understanding of aesthetic surgical procedures. After studying this article, the participant should be able to: 1. Explain the microbiology of Staphylococcus species and discuss antibiotic resistance development in Staphylococcus species and assess how clinical outcomes are affected. 2. Identify the epidemiology of Staphylococcus carriers and the impact on the clinical practice and regulation. Practice effective measures that prevent surgical site infections. 3. Practice screening for and decolonizing of patients with methicillin-resistant Staphylococcus aureus (MRSA). Physicians may earn 2.5 AMA PRA Category 1 Credit by successfully completing the examination based on material covered in this article. The examination begins on page 245. As a measure of the success of the education we hope you will receive from this article, we encourage you to log on to the Aesthetic Society website and take the preexamination before reading this article. Once you have completed the article, you may then take the examination again for CME credit. The Aesthetic Society will be able to compare your answers and use this data for future reference as we attempt to continually improve the CME articles we offer. ASAPS members can complete this CME examination online by logging on to the ASAPS Members-Only Website (http://www.surgery.org/members) and clicking on "Clinical Education" in the menu bar. Staphylococcus aureus is the most common cause of surgical site infections (SSI), with both methicillin-sensitive and methicillin-resistant strains causing these infections. The incidence of methicillin-resistant S aureus (MRSA) has increased in the US over the past decade, largely due to the emergence of community-acquired MRSA (CA-MRSA). This article reviews the microbiology and epidemiology of methicillin-sensitive S aureus (MSSA) and MRSA, risk factors for surgical site infections among plastic surgery patients, the evidence supporting preoperative screening and decolonization measures to prevent surgical site infections caused by MRSA, recommendations for anti-microbial prophylaxis, and treatment recommendations for surgical site infections. Other proven methods of reducing SSI, including maintenance of normothermia during surgery, glucose control, cessation of nicotine use, and not shaving the surgical site preoperatively are discussed.


A molecular dynamics study of oxygen ion diffusion in A-site ordered perovskite PrBaCo(2)O(5.5): data mining the oxygen trajectories.
Molecular dynamics (MD) simulations have been widely used to study oxygen ion diffusion in crystals. In the data analysis, one typically calculates the mean squared displacements to obtain the self-diffusion coefficients. Further information extraction for each individual atom poses significant challenges due to the lack of general methods. In this work, oxygen ion diffusion in A-site ordered perovskite PrBaCo2O5.5 is studied using MD simulations and the oxygen migration is analyzed by k-means clustering, a machine learning algorithm. The clustering analysis allows the tracking of each individual oxygen jump along with its corresponding location, i.e., oxygen site in BaO, PrO0.5 and CoO2 layers. Therefore it increases the understanding of the factors influencing oxygen diffusion. For example, it is found that the oxygen occupation fraction in the PrO0.5 layers increases with temperature, while in the CoO2 layers it decreases with temperature. Additionally, the activation enthalpies of oxygen jumps from CoO2 to CoO2, CoO2 to PrO0.5 and PrO0.5 to CoO2 are 0.22 eV, 0.54 eV and 0.34 eV, respectively, exhibiting anisotropic characteristics. Furthermore, the dwell times of oxygen atoms suggest that they are highly mobile in PrO0.5 layers. Combining the analysis of activation enthalpies and dwell times, it is suggested that the oxygen transport is fast within the CoO2 layers while the PrO0.5 layers work as oxygen vacancy reservoirs.


Complementary and Alternative Medicine Use in United States Adults With Liver Disease.
To describe the complementary and alternative medicine (CAM) use in US adults with liver disease. The prevalence and patterns of CAM use among US adults with liver disease have not been well characterized. The 2012 National Health Interview Survey is considered the most current and comprehensive source of information on CAM use in US adults. Using the results of the 2012 National Health Interview Survey, the prevalence of CAM use, most common modalities used, reasons for CAM use, perceived benefits, perceived helpfulness and importance, and disclosure of CAM to health care providers were compared between adults with and without liver disease. Of the 647 adults with liver disease, 41% reported using CAM in the prior year, compared with 33% of adults without liver disease. The most common modality was herbs and supplements (23%), and 3% of respondents reported consumption of a potentially hepatotoxic substance in the previous 30 days. Only a small proportion of CAM therapies were used specifically for liver disease, with milk thistle being the most common. Among respondents with liver disease, CAM was used more commonly for anxiety or depression, fatigue, and substance use. The majority believed that these therapies improved health. Nearly one-third of therapies were not reported to health care providers, mostly due to failure of the provider to ask. CAM use, particularly herbs and supplements, is prevalent among US adults with liver disease. Many do not disclose their CAM use to their providers, despite some using potentially hepatotoxic substances.


Ameliorative effect of astaxanthin on endothelial dysfunction in streptozotocin-induced diabetes in male rats.
The present study was designed to examine whether astaxanthin (ASX, 3,3-dihydroxybeta, beta-carotene-4,4-dione, CAS 472-61-7), a dietary antioxidant carotenoid that is naturally present in algae, crustaceans, and fish, has a protective effect on endothelial dysfunction of aortas in diabetic rats and the possible molecular mechanism involved. Male Wistar rats were randomly divided into four groups: control rats, diabetic rats, diabetic rats treated with ASX (10 mg/kg/d), and control rats treated with ASX. Type 1 diabetes was induced by a single intraperitoneal injection of streptozotocin (STZ; 60 mg/ kg). STZ-induced diabetes in rats was complicated with excessive oxidative stress and endothelial dysfunction, increased serum oxidized low-density lipoprotein (ox-LDL) and aortic malondialdehyde (MDA) levels, inhibited endothelium-dependent vasorelaxation to acetylcholine (ACh) and unaffected endothelium-dependent vasorelaxation to sodium nitroprusside (SNP). Simultaneously, lectin-like oxLDL receptor-i (LOX-1) expression was enhanced and endothelial nitric oxide (NO) synthase (eNOS) expression was reduced in the aortas of diabetic rats. ASX treatment could significantly decrease serum oxLDL and aortic MDA levels, attenuate blunted endothelium-dependent vasodilator responses to ACh, upregulate eNOS expression, and decrease LOX-1 expression. These results indicated that ASX could ameliorate diabetic endothelial dysfunction by inhibiting the ox-LDLLOX-1-eNOS pathway. Treatment with ASX might be clinically useful for diabetic complications associated with endothelial dysfunction.


RCAN1 regulates vesicle recycling and quantal release kinetics via effects on calcineurin activity.
We have previously shown that Regulator of Calcineurin 1 (RCAN1) regulates multiple stages of vesicle exocytosis. However, the mechanisms by which RCAN1 affects secretory vesicle exocytosis and quantal release kinetics remain unknown. Here, we use carbon fibre amperometry to detect exocytosis from chromaffin cells and identify these underlying mechanisms. We observe reduced exocytosis with repeated stimulations in chromaffin cells over-expressing RCAN1 (RCAN1(ox)), but not in wild-type (WT) cells, indicating a negative effect of RCAN1 on vesicle recycling and endocytosis. Acute exposure to calcineurin inhibitors, cyclosporine A and FK-506, replicates this effect in WT cells but has no additional effect in RCAN1(ox) cells. When we chronically expose WT cells to cyclosporine A and FK-506 we find that catecholamine release per vesicle and pre-spike foot (PSF) signal parameters are decreased, similar to that in RCAN1(ox) cells. Inhibiting calcineurin activity in RCAN1(ox) cells has no additional effect on the amount of catecholamine release per vesicle but further reduces PSF signal parameters. Although electron microscopy studies indicate these changes are not because of altered vesicle number or distribution in RCAN1(ox) cells, the smaller vesicle and dense core size we observe in RCAN1(ox) cells may underlie the reduced quantal release in these cells. Thus, our results indicate that RCAN1 most likely affects vesicle recycling and quantal release kinetics via the inhibition of calcineurin activity.


Effects of dietary supplementation of methionine and lysine on milk production and nitrogen utilization in dairy cows.
The effect of the content of lysine and methionine in metabolizable protein (MP) on lactation performance and N utilization in Chinese Holstein cows was determined. A control diet (C) was formulated to be adequate in energy but slightly limiting in MP. The concentration of Met and Lys in MP was 1.87 and 5.93%, respectively. The treatments were as follows (% of Met or Lys in MP): L=diet C supplemented with L-lysine-HCl at 0.49% on a dry matter (DM) basis (Met, 1.87; Lys, 7.00); M=diet C supplemented with 2-hydroxy-4-(methylthio)-butanoic acid (HMB) at 0.15% (Met, 2.35; Lys, 5.93); ML=diet C supplemented with 0.49% L-lysine HCl and 0.15% HMB (Met, 2.39; Lys, 7.10). The diets were fed to 60 Chinese Holsteins in mid-lactation (average days in milk=120, and milk yield=32.0 kg/d) for 8 wk. Milk yield was increased by supplementation of either Lys (1.5 kg/d) or Met (2.0 kg/d), and supplementation of both Lys and Met further increased milk yield (3.8 kg/d). There was no significant difference in dry matter intake across treatment groups. Cows on treatments M (3.95%) and ML (3.90%) had higher milk fat content than those on C (3.60%) and L (3.67%), but there were no significant differences in milk protein and lactose contents or somatic cell count among treatments. Supplementation of Met or Lys significantly increased Met or Lys concentration in arterial plasma. Treatment ML had a higher conversion of intake N to milk N and lower urea N concentrations in serum, urine, and milk than did treatment C. Supplementing HMB and L-lysine-HCl to provide approximately 2.3% Met and 7.0% Lys of the MP in diets slightly limiting in MP increased milk production, milk protein yield, and N utilization efficiency.


Relief of superior mesenteric artery syndrome with correction of multiplanar spinal deformity by posterior spinal fusion.
Superior mesenteric artery syndrome is obstruction of the third portion of the duodenum by compression between the abdominal aorta and superior mesenteric artery. Pediatric orthopedists are familiar with this entity, as the association between superior mesenteric artery syndrome and spinal fusion or body casting has been well established. However, patients with spinal deformities usually experience superior mesenteric artery syndrome after orthopedic intervention, with rates after corrective spinal surgery reported between 0.5% and 2.4%. Symptoms of superior mesenteric artery syndrome typically include nausea, bilious emesis, abdominal pain, early satiety, and anorexia. Initial treatment focuses on gastric decompression and maintaining euvolemia and electrolyte balance. The patient should receive enteral nutrition via nasojejunal tube or parenteral nutrition to allow for weight gain and subsequent resolution of the obstruction. The superior mesenteric artery takes off from the duodenum at an angle of 45 degrees to 60 degrees in normal individuals. The third portion of the duodenum is suspended between these vessels by the ligament of Treitz. Any variation in this relationship that decreases the arteriomesenteric angle may induce obstruction. Specifically, lumbar hyperextension or hyperlordosis can traction the mesentery and vessels. Only 2 cases of superior mesenteric artery syndrome in patients with sagittal plane spinal deformity have been described in the literature. In patients with concomitant superior mesenteric artery syndrome and spinal deformity, correction of the deformity may help alleviate the obstruction and result in faster recovery. The contribution of spinal column deformity to the arteriomesenteric angle should not be overlooked.


Differential impact of P-glycoprotein (ABCB1) and breast cancer resistance protein (ABCG2) on axitinib brain accumulation and oral plasma pharmacokinetics.
The second-generation tyrosine kinase inhibitor and anticancer drug axitinib is a potent, orally active inhibitor of the vascular endothelial growth factor receptors 1, 2, and 3. Axitinib has clinical activity against solid tumors such as metastatic renal cell carcinoma and advanced pancreatic cancer. We studied axitinib transport using Madin-Darby canine kidney II cells overexpressing human ABCB1 or ABCG2 or murine Abcg2. Axitinib was a good substrate of ABCB1 and Abcg2, whereas transport activity by ABCG2 was moderate. These transporters may therefore contribute to axitinib resistance in tumor cells. Upon oral administration of axitinib, Abcg2(-/-) and Abcb1a/1b;Abcg2(-/-) mice displayed 1.7- and 1.8-fold increased axitinib areas under the plasma concentration-time curve from 0 to 4 compared with those of wild-type mice. Plasma concentrations in Abcb1a/1b(-/-) mice were not significantly increased. In contrast, relative brain accumulation of axitinib in Abcb1a/1b(-/-) and Abcb1a/1b;Abcg2(-/-) mice was, respectively, 6.8- and 13.9-fold higher than that in wild-type mice at 1 h and 4.9- and 20.7-fold at 4 h after axitinib administration. In Abcg2(-/-) mice, we found no significant differences in brain accumulation compared with those in wild-type mice. Thus, Abcb1 strongly restricts axitinib brain accumulation and completely compensates for the loss of Abcg2 at the blood-brain barrier, whereas Abcg2 can only partially take over Abcb1-mediated axitinib efflux. Hence, Abcg2 has a stronger impact on axitinib oral plasma pharmacokinetics, whereas Abcb1 is the more important transporter at the blood-brain barrier. These findings illustrate that in vitro transport data for ABCB1 and ABCG2 cannot always be simply extrapolated to the prediction of the relative impact of these transporters on oral availability versus brain penetration.


Apnoeic episodes induced by smothering: two cases identified by covert video surveillance.
Recurrent cyanotic episodes associated on some occasions with loss of consciousness due to cerebral hypoxia were investigated by long term tape recordings of breathing activity, oxygen saturation, air flow, electrocardiographic activity, and in some cases electroencephalographic activity. In 51 infants and children the mechanisms for the cyanotic episodes were identified (prolonged expiratory apnoea in 45, sleep related airway obstruction in three, seizure induced apnoea in one, behaviour induced apnoea in one). In one child apnoea was suspected as being caused by suffocation (smothering) by the mother. This was confirmed after enlisting the help of the police, who undertook covert video surveillance during cyanotic episodes. Each cyanotic episode was associated with a pattern of disturbance on the multichannel tape recordings which may be pathognomonic of this type of apnoea. A second infant with cyanotic episodes in whom smothering was suspected was referred for similar investigation after the availability of video recordings became established. Maternal smothering was again supported by specific patterns on multichannel tape recordings and confirmed by video surveillance. Diagnosis by video surveillance produces unequivocal evidence in these cases and avoids the need for medical and nursing staff to confront the mother with a possibly incorrect suspicion or in a court of law.


Investigation of interocular blur suppression using luminance-modulated and contrast-modulated noise stimuli.
Presenting two sufficiently dissimilar images, one to each eye, may result in interocular suppression. The present study measured interocular suppression depth and extent in binocularly normal participants when blurring one eye only with varying dioptric lens powers (+0.5, +1, +2, and +4 D). Visual stimuli consisted of eight concentric rings of alternate polarity, divisible into eight sectors, within the central circular 24° visual field. Binocular "ring" stimuli therefore consisted of 64 individually testable dichoptic sectors. Using a two-alternative forced choice paradigm with a staircase procedure, signal strength of each dichoptic sector in the blurred eye was adjusted to perceptually match that of the surrounding ring from the nonblurred eye, determining the point of subjective equality. Rings were defined by differences in luminance (L), luminance-modulated noise (LM), or contrast-modulated noise (CM). Suppression depth was similar irrespective of sector location within the visual field and increased with increasing difference in interocular blur. Adding dynamic noise (LM vs. L stimuli) reduced the effect of blur on measured suppression depth. Significantly deeper suppression was measured for CM than for LM stimuli, both created using dynamic noise, the difference increasing at higher levels of interocular blur. As binocularity is disrupted with interocular blur, this result suggests that CM envelope combination may be processed by later mechanisms receiving binocular input than those required for the processing of LM stimuli. Differences in suppression depth between LM and CM stimuli could not be attributed to differences in spatial summation properties, stimulus visibility, noise modulation, or differential effects on blur discriminability.


TLR3 drives IRF6-dependent IL-23p19 expression and p19/EBI3 heterodimer formation in keratinocytes.
Interferon regulatory factor (IRF) family members impart cell-type specificity to toll-like receptor (TLR) signalling, and we recently identified a role for IRF6 in TLR2 signalling in epithelial cells. TLR3 has a well-characterized role in wound healing in the skin, and here, we examined TLR3-dependent IRF6 functions in human keratinocytes. Primary keratinocytes responded robustly to the TLR3 agonist poly(IC) with upregulation of mRNAs for interferon-β (IFN-β), the interleukin-12 (IL-12) family member IL-23p19 and the chemokines IL-8 and chemokine (C-C motif) ligand 5 (CCL5). Silencing of IRF6 expression enhanced poly(IC)-inducible IFN-β mRNA levels and inhibited poly(IC)-inducible IL-23p19 mRNA expression in primary keratinocytes. Consistent with these data, co-transfection of IRF6 increased poly(IC)-inducible IL-23p19 promoter activity, but inhibited poly(IC)-inducible IFN-β promoter activity in reporter assays. Surprisingly, poly(IC) did not regulate IL-12p40 expression in keratinocytes, suggesting that TLR3-inducible IL-23p19 may have an IL-23-independent function in these cells. The only other IL-12 family member that was strongly poly(IC) inducible was EBI3, which has not been shown to heterodimerize with IL-23p19. Both co-immunoprecipitation and proximity ligation assays revealed that IL-23p19 and EBI3 interact in cells. Co-expression of IL-23p19 and EBI3, as compared with IL-23p19 alone, resulted in increased levels of secreted IL-23p19, implying a functional role for this heterodimer. In summary, we report that IRF6 regulates a subset of TLR3 responses in human keratinocytes, including the production of a novel IL-12 family heterodimer (p19/EBI3). We propose that the TLR3-IRF6-p19/EBI3 axis may regulate keratinocyte and/or immune cell functions in the context of cell damage and wound healing in the skin.


Focus issue: fine-tuning Hedgehog signaling in development and disease.
Hedgehog (Hh) signaling plays varied and critical roles in animal development, homeostasis, and disease. The core components of the canonical pathway and many key modulators of signaling have been identified, as have mechanisms by which the quantity and quality of signaling through the pathway are modulated. New research reveals details about generation and interpretation of Hh gradients, noncanonical Hh signaling, and fine-tuning the biological response to Hh signaling, particularly in the context of human diseases such as cancer.


Genomic distribution and estimation of nucleotide diversity in natural populations: perspectives from the collared flycatcher (Ficedula albicollis) genome.
Properly estimating genetic diversity in populations of nonmodel species requires a basic understanding of how diversity is distributed across the genome and among individuals. To this end, we analysed whole-genome resequencing data from 20 collared flycatchers (genome size ≈1.1 Gb; 10.13 million single nucleotide polymorphisms detected). Genomewide nucleotide diversity was almost identical among individuals (mean = 0.00394, range = 0.00384-0.00401), but diversity levels varied extensively across the genome (95% confidence interval for 200-kb windows = 0.0013-0.0053). Diversity was related to selective constraint such that in comparison with intergenic DNA, diversity at fourfold degenerate sites was reduced to 85%, 3' UTRs to 82%, 5' UTRs to 70% and nondegenerate sites to 12%. There was a strong positive correlation between diversity and chromosome size, probably driven by a higher density of targets for selection on smaller chromosomes increasing the diversity-reducing effect of linked selection. Simulations exploring the ability of sequence data from a small number of genetic markers to capture the observed diversity clearly demonstrated that diversity estimation from finite sampling of such data is bound to be associated with large confidence intervals. Nevertheless, we show that precision in diversity estimation in large outbred population benefits from increasing the number of loci rather than the number of individuals. Simulations mimicking RAD sequencing showed that this approach gives accurate estimates of genomewide diversity. Based on the patterns of observed diversity and the performed simulations, we provide broad recommendations for how genetic diversity should be estimated in natural populations.


Frequency and Impact of Cerebral Infarctions in Patients With Tuberculous Meningitis.
Background and Purpose- Cerebral infarctions complicate a variable proportion of tuberculous meningitis (TBM) cases and adversely affect outcomes. The objective of this study was to evaluate the predictors of cerebral infarcts in patients with TBM and to assess their impact on mortality. Methods- The study was based on a retrospective chart review of all patients with TBM admitted to a tertiary care hospital between 2002 and 2013. Data were collected on basic demographics, conventional vascular risk factors, radiological findings, severity of TBM, and neurological outcomes. Data were analyzed using SPSS version 19.0. Binary logistic regression was done to determine the factors predictive of cerebral infarcts and of mortality in patients with TBM. Results- A total of 559 patients were admitted with TBM during the study period. Mean age was 41.9 years (SD, 17.7 years), and 47% were women. A quarter of the patients had stage III disease. One hundred forty-four (25.8%) patients had cerebral infarcts on brain imaging of which 3 quarters were acute or subacute. Those with cerebral infarcts were more likely to be >40 years of age (adjusted odds ratio [AOR], 1.7; 95% CI, 1.1-2.7) and to have hypertension (AOR, 1.8; 95% CI, 1.1-2.8), dyslipidemia (AOR, 9.7; 95% CI, 3.8-24.8), and diabetes mellitus (AOR, 2.2; 95% CI, 1.3-3.6). Presence of cerebral infarction was an independent predictor of mortality among patients with TBM (AOR, 2.1; 95% CI, 1.22-3.5). Conclusions- Cerebral infarcts complicate a substantial proportion of TBM cases. Conventional vascular risk factors are the most important predictors of infarction, and future efforts need to focus on these high-risk patients with TBM to reduce morbidity and mortality.


Lack of Skeletal Muscle IL-6 Affects Pyruvate Dehydrogenase Activity at Rest and during Prolonged Exercise.
Pyruvate dehydrogenase (PDH) plays a key role in the regulation of skeletal muscle substrate utilization. IL-6 is produced in skeletal muscle during exercise in a duration dependent manner and has been reported to increase whole body fatty acid oxidation, muscle glucose uptake and decrease PDHa activity in skeletal muscle of fed mice. The aim of the present study was to examine whether muscle IL-6 contributes to exercise-induced PDH regulation in skeletal muscle. Skeletal muscle-specific IL-6 knockout (IL-6 MKO) mice and floxed littermate controls (control) completed a single bout of treadmill exercise for 10, 60 or 120 min, with rested mice of each genotype serving as basal controls. The respiratory exchange ratio (RER) was overall higher (P<0.05) in IL-6 MKO than control mice during the 120 min of treadmill exercise, while RER decreased during exercise independent of genotype. AMPK and ACC phosphorylation also increased with exercise independent of genotype. PDHa activity was in control mice higher (P<0.05) at 10 and 60 min of exercise than at rest but remained unchanged in IL-6 MKO mice. In addition, PDHa activity was higher (P<0.05) in IL-6 MKO than control mice at rest and 60 min of exercise. Neither PDH phosphorylation nor acetylation could explain the genotype differences in PDHa activity. Together, this provides evidence that skeletal muscle IL-6 contributes to the regulation of PDH at rest and during prolonged exercise and suggests that muscle IL-6 normally dampens carbohydrate utilization during prolonged exercise via effects on PDH.


Evaluation of aortic intima-media thickness in newborns with Down syndrome.
Health care programs for children with Down syndrome (DS) help improve the overall outcome and quality of life of these children. It is therefore very important to focus on the most common and serious problems of these patients, such as congenital heart defects and cardiac problems, and to keep medical guidelines updated with regard to these problems. The aim of this study was to evaluate aortic intima-media thickness (aIMT), lipid profiles and blood pressure in DS patients in comparison with a control group of ageand gender-matched neonates without DS. Serum concentrations of lipids (total cholesterol, LDL-cholesterol, HDL-cholesterol, and triglyceride), blood pressure and abdominal aIMT were measured in newborns with DS, and compared with the measurements from ageand gender-matched newborns without DS. No statistically significant differences between the 2 groups of newborns were detected with respect to aIMT, lipid levels or blood pressure. This study represents the first investigation of aIMT - one of the most important indicators of atherosclerosis - in DS patients, but neither a significantly increased aIMT, nor any significant changes in lipid profiles or blood pressure were detected in this group of patients. Whether aIMT differs according to the type of congenital heart defect (such as atrial septal defect, ventricular septal defect, patent ductus arteriosus, and atrioventricular septal defect) among DS patients remains to be determined in future studies.


Transcranial Doppler detection of anterior cerebral artery vasospasm.
The performance of transcranial Doppler in the detection of anterior cerebral artery vasospasm and vasospasm in patients after subarachnoid haemorrhage was analysed. Transcranial Doppler and cerebral angiography were performed within the same 24 hours on each of 41 patients with acute subarachnoid haemorrhage. Sensitivity and specificity of transcranial Doppler to classify middle cerebral arteries, anterior cerebral arteries, and patients with angiographic vasospasm were determined at mean velocities of 120 and 140 cm/s. Accuracy of transcranial Doppler was better at 140 than at 120 cm/s. For the middle cerebral artery, sensitivity was 86%, specificity 98%. For the anterior cerebral artery, sensitivity was 13%, specificity 100%. Among all patients, sensitivity was 45%, specificity 96%. Among patients with anterior communicating artery aneurysms, sensitivity was 14%, specificity 90%. Therefore, transcranial Doppler accurately differentiates between middle cerebral arteries with and without vasospasm on angiography, but has a very low sensitivity for detecting anterior cerebral artery vasospasm and vasospasm in patients with anterior communicating artery aneurysms. Since vasospasm may involve anterior cerebral arteries while sparing middle cerebral arteries, especially after rupture of an anterior communicating artery aneurysm, caution should be exercised in using negative transcranial Doppler results to make treatment decisions based on the assumed absence of vasospasm.


Double mitral valve orifice in atrioventricular defects.
Eleven patients with double mitral valve orifice and atrioventricular defects were studied, and the diagnosis proven by open heart surgery. The correct preoperative diagnosis was suggested by a characteristic angiographic appearance of the medial border of the left ventricle. M-mode echocardiography may show the two orifices which are better seen on two dimensional echocardiography. Two operative deaths occurred in patients with a slightly obstructive valve, and one late death followed mitral valve replacement. Eight patients did well after minor repair to a cleft, or leaving the valve untouched. Occasionally a double mitral orifice, if disorganised and slightly stenotic, may cause early deterioration in a patient with an otherwise uncomplicated ostium primum defect. The surgeon should be forewarned of this mitral anomaly since simple repair may be difficult.


Resisting the screening imperative: patienthood, populations and politics in prostate cancer detection technologies for the UK.
The introduction of mass screening programmes in the UK has been controversial. It is instructive to examine medical conditions for which screening has been actively considered but not introduced, such as prostate cancer. Incidence of the disease has escalated during the last 20 years, partly due to the upsurge in use of PSA (prostate-specific antigen) detection technology. The controversy is moving into a new phase, associated with the development of new molecular genetic biomarkers and tests derived from genome-association studies. The paper outlines the most recent scientific and technological developments for the three types of detection technology - PSA, genetic, and genomic. Applying concepts of risk, technology governance and technology expectations, it is shown that central public health governance actors continue to resist the tidal wave of new technologies through a variety of increasingly diverse governance modes. In the case of PSA, a governance trend moving beyond 'responsibilisation' to citizen rights is shown, whereas in the case of genetic tests and genomic risk profiling, state public health agencies are shown to be engaging in a form of technology expectation management, as it responds to a new marketplace of private commercial testing and mediatised science-based visions of future healthcare.


Development of an imprinted polymeric sensor with dual sensing property for trace level estimation of zinc and arginine.
Using virtuousness of multi-template imprinting and selectivity of single-template imprinting, for the first time, we have prepared an imprinted polymer matrix that can detect two target analyte viz., Zn(2+) and arginine in single motif. For the preparation of imprinted polymer network, a new monomer {2-acrylamido-4-(methylthio)butanoic acid} was synthesized, which gives a biocompatible polymer network. Herein, the synthesized imprinted polymer mixed with multi-walled carbon nanotubes was immobilized onto the surface of platinum electrode to develop an electrochemical sensor for quantitative recognition of Zn(2+) and arginine using differential pulse voltammetry. Cyclic voltammetry, electrochemical impedance spectroscopy, and chronocoulometric analysis were also performed to study the kinetics of zinc-arginine complex. Under the optimal conditions the linear range of the calibration curve for Zn(2+) and arginine was 0.04-310.0ngmL(-1) and 0.04-235.0ngmL(-1), with the detection limit of 18.0 and 15.0pgmL(-1). The sensor has been successfully applied to the determination of Zn(2+) and arginine in water, food, and pharmaceutical samples and achieved high sensitivity and selectivity.


Influence of serotonergic mechanisms on the urine flow rate in male rats.
This study extensively examined the role of a 5-HT(1A) receptor in controlling voiding function in anesthetized male rats. A simultaneous recording of the intravesical pressure (IVP), external urethral sphincter (EUS)-electromyography (EMG), and urine flow rate (UFR) during continuous cystometry was used. 8-Hydroxy-2-(di-n-propylamino)-tetralin (8-OH-DPAT), a 5-HT(1A) receptor agonist, significantly improved the voiding efficiency, as detected by increases in the evoked contraction amplitude, EUS burst period, and silent period, and decreases in the volume threshold, pressure threshold, and residual volume. Interestingly, the UFR during voiding was reduced by 8-OH-DPAT, as evidenced by decreases in the maximal UFR and mean UFRs of the voiding period, spike duration, and interspike interval. Conversely, treating rats with WAY-100635, a 5-HT(1A) antagonist, produced effects opposite to those produced by 8-OH-DPAT. These findings suggest that 8-OH-DPAT improved the voiding efficiency by enhancing the detrusor contractile ability and prolonging EUS burst period, which would compensate for the lower UFR, resulting from urethral smooth muscle contractions and longer EUS silent periods during voiding. The present study contributes to our understanding of the role of 5-HT(1A) receptors in controlling the urine flow rate in male rats.


Large carbon isotope fractionation associated with oxidation of methyl halides by methylotrophic bacteria.
The largest biological fractionations of stable carbon isotopes observed in nature occur during production of methane by methanogenic archaea. These fractionations result in substantial (as much as approximately 70 per thousand) shifts in delta(13)C relative to the initial substrate. We now report that a stable carbon isotopic fractionation of comparable magnitude (up to 70 per thousand) occurs during oxidation of methyl halides by methylotrophic bacteria. We have demonstrated biological fractionation with whole cells of three methylotrophs (strain IMB-1, strain CC495, and strain MB2) and, to a lesser extent, with the purified cobalamin-dependent methyltransferase enzyme obtained from strain CC495. Thus, the genetic similarities recently reported between methylotrophs, and methanogens with respect to their pathways for C(1)-unit metabolism are also reflected in the carbon isotopic fractionations achieved by these organisms. We found that only part of the observed fractionation of carbon isotopes could be accounted for by the activity of the corrinoid methyltransferase enzyme, suggesting fractionation by enzymes further along the degradation pathway. These observations are of potential biogeochemical significance in the application of stable carbon isotope ratios to constrain the tropospheric budgets for the ozone-depleting halocarbons, methyl bromide and methyl chloride.


Long-term survival and healthcare utilization outcomes attributable to sepsis and pneumonia.
Hospital associated infections are major problems, which are increasing in incidence and very costly. However, most research has focused only on measuring consequences associated with the initial hospitalization. We explored the long-term consequences of infections in elderly Medicare patients admitted to an intensive care unit (ICU) and discharged alive, focusing on: sepsis, pneumonia, central-line-associated bloodstream infections (CLABSI), and ventilator-associated pneumonia (VAP); the relationships between the infections and long-term survival and resource utilization; and how resource utilization was related to impending death during the follow up period. Clinical data and one year pre- and five years post-index hospitalization Medicare records were examined. Hazard ratios (HR) and healthcare utilization incidence ratios (IR) were estimated from state of the art econometric models. Patient demographics (i.e., age, gender, race and health status) and Medicaid status (i.e., dual eligibility) were controlled for in these models. In 17,537 patients, there were 1,062 sepsis, 1,802 pneumonia, 42 CLABSI and 52 VAP cases. These subjects accounted for 62,554 person-years post discharge. The sepsis and CLABSI cohorts were similar as were the pneumonia and VAP cohorts. Infection was associated with increased mortality (sepsis HR = 1.39, P < 0.01; and pneumonia HR = 1.58, P < 0.01) and the risk persisted throughout the follow-up period. Persons with sepsis and pneumonia experienced higher utilization than controls (e.g., IR for long-term care utilization for those with sepsis ranged from 2.67 to 1.93 in years 1 through 5); and, utilization was partially related to impending death. The infections had significant and lasting adverse consequences among the elderly. Yet, many of these infections may be preventable. Investments in infection prevention interventions are needed in both community and hospitals settings.


Teaching ethical analysis in occupational therapy.
Ethical decision making is a cognitive skill requiring education in ethical principles and an understanding of specific ethical issues. It is also a psychodynamic process involving personalities, values, opinions, and perceptions. This article proposes the use of case studies and role-playing techniques in teaching ethics in occupational therapy to supplement conventional methods of presenting ethical theories and principles. These two approaches invite students to discuss and analyze crucial issues in occupational therapy from a variety of viewpoints. Methodology of developing case studies and role-playing exercises are discussed. The techniques are evaluated and their application to the teaching of ethics is examined.


Cardiac arrest and sudden death in dialysis units.
For patients with end-stage renal disease and their providers, dialysis unit-based cardiac arrest is the most feared complication of hemodialysis. However, relatively little is known regarding its frequency or epidemiology, or whether a fraction of these events could be prevented. To explore clinical correlates of dialysis unit-based cardiac arrest, 400 reported arrests over a nine-month period from October 1998 through June 1999 were reviewed in detail. Clinical characteristics of patients who suffered cardiac arrest were compared with a nationally representative cohort of> 77,000 hemodialysis patients dialyzed at Fresenius Medical Care North America-affiliated facilities. The cardiac arrest rate was 400 out of 5,744,708, corresponding to a rate of 7 per 100,000 hemodialysis sessions. Cardiac arrest was more frequent during Monday dialysis sessions than on other days of the week. Case patients were nearly twice as likely to have been dialyzed against a 0 or 1.0 mEq/L potassium dialysate on the day of cardiac arrest (17.1 vs. 8.8%). Patients who suffered a cardiac arrest were on average older (66.3 +/- 12.9 vs. 60.2 +/- 15.4 years), more likely to have diabetes (61.8 vs. 46.8%), and more likely to use a catheter for vascular access (34.1 vs. 27.8%) than the general hemodialysis population. Sixteen percent of patients experienced a drop in systolic pressure of 30 mm Hg or more prior to the arrest. Thirty-seven percent of patients who suffered cardiac arrest had been hospitalized within the past 30 days. Sixty percent of patients died within 48 hours of the arrest, including 13% while in the dialysis unit. Cardiac arrest is a relatively infrequent but devastating complication of hemodialysis. To reduce the risk of adverse cardiac events on hemodialysis, the dialysate prescription should be evaluated and modified on an ongoing basis, especially following hospitalization in high-risk patients.


Adenosine N1-Oxide Exerts Anti-inflammatory Effects through the PI3K/Akt/GSK-3β Signaling Pathway and Promotes Osteogenic and Adipocyte Differentiation.
Previously, we reported that adenosine N1-oxide (ANO), which is found in royal jelly, inhibited the secretion of inflammatory mediators by activated macrophages and reduced lethality in lipopolysaccharide (LPS)-induced endotoxin shock. Here, we examined the regulatory mechanisms of ANO on the release of pro-inflammatory cytokines, with a focus on the signaling pathways activated by toll-like receptor (TLR)4 in response to LPS. ANO inhibited both tumor necrosis factor (TNF)-α and interleukin (IL)-6 secretion from LPS-stimulated RAW264.7 cells without affecting cell proliferation. In this response, phosphorylation of mitogen-activated protein kinase (MAPK) family members (extracellular signal-regulated kinase (ERK)1/2, p38 and SAPK/c-Jun N-terminal kinase (JNK)) and nuclear factor-κB (NF-κB) p65 was not affected by treatment with ANO. In contrast, phosphorylation of Akt (Ser473) and its downstream molecule glycogen synthase kinase-3β (GSK-3β) (Ser9) was up-regulated by ANO, suggesting that ANO stimulated GSK-3β phosphorylation via phosphatidylinositol 3-kinase (PI3K)/Akt signaling pathway. The phosphorylation of GSK-3β on Ser9 has been shown to negatively regulate the LPS-induced inflammatory response. Activation of PI3K/Akt signaling pathway has also been implicated in differentiation of mesenchymal stem cells into osteoblasts and adipocytes. As expected, ANO induced alkaline phosphatase activity and promoted calcium deposition in a mouse pre-osteoblastic MC3T3-E1 cell line. The ANO-induced differentiation into osteoblasts was abrogated by coincubation with Wortmannin. Furthermore, ANO promoted insulin/dexamethasone-induced differentiation of mouse 3T3-L1 preadipocytes into adipocytes at much lower concentrations than adenosine. The protective roles of PI3K/Akt/GSK-3β signaling pathway in inflammatory disorders have been well documented. Our data suggest that ANO may serve as a potential candidate for the treatment of inflammatory disorders. Promotion of osteogenic and adipocyte differentiation further suggests its application for regenerative medicine.


Severity of bias of a simple estimator of the causal odds ratio in Mendelian randomization studies.
Mendelian randomization studies estimate causal effects using genetic variants as instruments. Instrumental variable methods are straightforward for linear models, but epidemiologists often use odds ratios to quantify effects. Also, odds ratios are often the quantities reported in meta-analyses. Many applications of Mendelian randomization dichotomize genotype and estimate the population causal log odds ratio for unit increase in exposure by dividing the genotype-disease log odds ratio by the difference in mean exposure between genotypes. This 'Wald-type' estimator is biased even in large samples, but whether the magnitude of bias is of practical importance is unclear. We study the large-sample bias of this estimator in a simple model with a continuous normally distributed exposure, a single unobserved confounder that is not an effect modifier, and interpretable parameters. We focus on parameter values that reflect scenarios in which we apply Mendelian randomization, including realistic values for the degree of confounding and strength of the causal effect. We evaluate this estimator and the causal odds ratio using numerical integration and obtain approximate analytic expressions to check results and gain insight. A small simulation study examines finite sample bias and mild violations of the normality assumption. For our simple data-generating model, we find that the Wald estimator is asymptotically biased with a bias of around 10% in fairly typical Mendelian randomization scenarios but which can be larger in more extreme situations. Recently developed methods such as structural mean models require fewer untestable assumptions and we recommend their use when the individual-level data they require are available. The Wald-type estimator may retain a role as an approximate method for meta-analysis based on summary data.


"The biggest problem we've ever had to face": how families manage driving cessation with people with dementia.
It is recognized that people with dementia are likely to need to stop driving at some point following diagnosis. Driving cessation can lead to negative outcomes for people with dementia and their family caregivers (FC), who often experience family conflict and tension throughout the process. Family experiences surrounding driving cessation have begun to be explored but warrant further examination. Using a descriptive phenomenological approach, semi-structured interviews were undertaken with key stakeholders, including 5 retired drivers with dementia, 12 FC, and 15 health professionals (HP). Data were analyzed inductively to explore the needs and experiences of people with dementia and FC. The data revealed a range of possible interactions between people with dementia and FC. These were organized into a continuum of family dynamics according to levels of collaboration and conflict: in it together, behind the scenes, active negotiations, and at odds. At the in it together end of the continuum, people with dementia and FC demonstrated collaborative approaches and minimal conflict in managing driving cessation. At the at odds end, they experienced open conflict and significant tension in their interactions. Contextual factors influencing family dynamics were identified, along with the need for individualized approaches to support. The continuum of family dynamics experienced during driving cessation may help clinicians better understand and respond to complex family needs. Interventions should be tailored to families' distinctive needs with consideration of their unique contextual factors influencing dynamics, to provide sensitive and responsive support for families managing driving cessation.


The development and initial validation of a clinical tool for patients' preferences on patient participation--The 4Ps.
To report on the development and initial testing of a clinical tool, The Patient Preferences for Patient Participation tool (The 4Ps), which will allow patients to depict, prioritize, and evaluate their participation in health care. While patient participation is vital for high quality health care, a common definition incorporating all stakeholders' experience is pending. In order to support participation in health care, a tool for determining patients' preferences on participation is proposed, including opportunities to evaluate participation while considering patient preferences. Exploratory mixed methods studies informed the development of the tool, and descriptive design guided its initial testing. The 4Ps tool was tested with 21 Swedish researcher experts (REs) and patient experts (PEs) with experience of patient participation. Individual Think Aloud interviews were employed to capture experiences of content, response process, and acceptability. 'The 4Ps' included three sections for the patient to depict, prioritize, and evaluate participation using 12 items corresponding to 'Having Dialogue', 'Sharing Knowledge', 'Planning', and 'Managing Self-care'. The REs and PEs considered 'The 4Ps' comprehensible, and that all items corresponded to the concept of patient participation. The tool was perceived to facilitate patient participation whilst requiring amendments to content and layout. A tool like The 4Ps provides opportunities for patients to depict participation, and thus supports communication and collaboration. Further patient evaluation is needed to understand the conditions for patient participation. While The 4Ps is promising, revision and testing in clinical practice is required.


Establishing glucose- and ABA-regulated transcription networks in Arabidopsis by microarray analysis and promoter classification using a Relevance Vector Machine.
Establishing transcriptional regulatory networks by analysis of gene expression data and promoter sequences shows great promise. We developed a novel promoter classification method using a Relevance Vector Machine (RVM) and Bayesian statistical principles to identify discriminatory features in the promoter sequences of genes that can correctly classify transcriptional responses. The method was applied to microarray data obtained from Arabidopsis seedlings treated with glucose or abscisic acid (ABA). Of those genes showing >2.5-fold changes in expression level, approximately 70% were correctly predicted as being up- or down-regulated (under 10-fold cross-validation), based on the presence or absence of a small set of discriminative promoter motifs. Many of these motifs have known regulatory functions in sugar- and ABA-mediated gene expression. One promoter motif that was not known to be involved in glucose-responsive gene expression was identified as the strongest classifier of glucose-up-regulated gene expression. We show it confers glucose-responsive gene expression in conjunction with another promoter motif, thus validating the classification method. We were able to establish a detailed model of glucose and ABA transcriptional regulatory networks and their interactions, which will help us to understand the mechanisms linking metabolism with growth in Arabidopsis. This study shows that machine learning strategies coupled to Bayesian statistical methods hold significant promise for identifying functionally significant promoter sequences.


Seasonal variations in cardiovascular disease.
Cardiovascular disease (CVD) follows a seasonal pattern in many populations. Broadly defined winter peaks and clusters of all subtypes of CVD after 'cold snaps' are consistently described, with corollary peaks linked to heat waves. Individuals living in milder climates might be more vulnerable to seasonality. Although seasonal variation in CVD is largely driven by predictable changes in weather conditions, a complex interaction between ambient environmental conditions and the individual is evident. Behavioural and physiological responses to seasonal change modulate susceptibility to cardiovascular seasonality. The heterogeneity in environmental conditions and population dynamics across the globe means that a definitive study of this complex phenomenon is unlikely. However, given the size of the problem and a range of possible targets to reduce seasonal provocation of CVD in vulnerable individuals, scope exists for both greater recognition of the problem and application of multifaceted interventions to attenuate its effects. In this Review, we identify the physiological and environmental factors that contribute to seasonality in nearly all forms of CVD, highlight findings from large-scale population studies of this phenomenon across the globe, and describe the potential strategies that might attenuate peaks in cardiovascular events during cold and hot periods of the year.


The endocannabinoid system, cannabinoids, and pain.
The endocannabinoid system is involved in a host of homeostatic and physiologic functions, including modulation of pain and inflammation. The specific roles of currently identified endocannabinoids that act as ligands at endogenous cannabinoid receptors within the central nervous system (primarily but not exclusively CB 1 receptors) and in the periphery (primarily but not exclusively CB 2 receptors) are only partially elucidated, but they do exert an influence on nociception. Exogenous plant-based cannabinoids (phytocannabinoids) and chemically related compounds, like the terpenes, commonly found in many foods, have been found to exert significant analgesic effects in various chronic pain conditions. Currently, the use of Δ9-tetrahydrocannabinol is limited by its psychoactive effects and predominant delivery route (smoking), as well as regulatory or legal constraints. However, other phytocannabinoids in combination, especially cannabidiol and β-caryophyllene, delivered by the oral route appear to be promising candidates for the treatment of chronic pain due to their high safety and low adverse effects profiles. This review will provide the reader with the foundational basic and clinical science linking the endocannabinoid system and the phytocannabinoids with their potentially therapeutic role in the management of chronic pain.


Amino acid containing thapsigargin analogues deplete androgen receptor protein via synthesis inhibition and induce the death of prostate cancer cells.
There are quantitative and/or qualitative mechanisms allowing androgen receptor (AR) growth signaling in androgen ablation refractory prostate cancer cells. Regardless of the mechanism, agents that deplete AR protein expression prevent such AR growth signaling. Thapsigargin (TG) is a highly cell-penetrant sequiterpene-lactone that once inside cells inhibits (IC(50), ∼ 10 nmol/L) critically important housekeeping SERCA 2b calcium pumps in the endoplasmic reticulum. Using a series of five genetically diverse androgen ablation refractory human prostate cancer lines (LNCaP, LAPC-4, VCaP, MDA-PCa-2b, and CWR22Rv1), TG inhibition of SERCA pumps consistently results in depletion of the endoplasmic reticulum Ca(+2) coupled with μmol/L elevation in the intracellular free Ca(+2) initiating a molecular cascade that: (a) inhibits Cap-dependent AR protein synthesis resulting in 90% depletion of AR protein by 24 hours of TG exposure, (b) arrests the cells in G(0), and (c) induces their apoptotic death. Unfortunately, due to its highly lipophilic nature, TG is not deliverable as a systemic agent without host toxicity. Therefore, TG analogues containing amino acids were developed, which retain ability to deplete AR protein and induce cell death and which can be covalently linked to peptide carriers producing water soluble prodrugs for systemic delivery. Specific amino acid sequences are used to restrict the liberation of cytotoxic amino acid containing TG analogues from the peptide prodrug by prostate-specific proteases, such as prostate-specific antigen and prostate-specific membrane antigen, or cancer-specific proteases, such as fibroblast activation protein, so that toxicity of these prodrugs is selectively targeted to metastatic sites of prostate cancer. Based on these results, these prodrugs are undergoing clinical development.


Evaluation of the placental environment with a new in vitro model of histocultures of early and term placentae: determination of cytokine and chemokine expression profiles.
We aimed to set up and validate a new in vitro model of placental histocultures, for the evaluation of cytokine and chemokine profiles of the placental environment, over a long culture period. Micro-explant cultures from 6 early and 6 term placentae were set up on collagen sponge gel supports at a liquid/air interface. At various times during culture, we analyzed tissue morphology and cell death by microscopy and quantified beta-hCG production and mRNA levels for beta-hCG and insulin-like 4 (INSL4). Levels of IL-6, LIF, TNF alpha, IL-10, IFN-gamma, IL-16 and RANTES in the medium were measured by ELISA on days 1, 4 and 7 of culture. SDF-1 mRNA expression was determined by real-time PCR at the same time points. Histocultures from early and term placentae remained viable until day 10. High levels of IL-6 and LIF production, low levels of TNF alpha, IL-10 and IFN-gamma production and significant SDF-1 expression were observed. These data indicate that placental histoculture is a suitable and reliable in vitro model for studying the placental environment.


Nineteenth century seeds reveal the population genetics of landrace barley (Hordeum vulgare).
Barley (Hordeum vulgare) is a major crop, grown worldwide and in a wide range of climatic conditions. Despite its importance as a crop species, little is known about the population genetics of barley and the effects of bottlenecks, adaptation, and gene flow on genetic diversity within and between landrace populations. In areas with highly developed agriculture, such as Northern Europe, these types of genetic studies are hampered by lack of landraces preserved in situ or ex situ. Here, we report a genetic study of Swedish landrace barley using 113-year-old seed samples. The results demonstrate differing levels of variation with some latitudinal effect. We also detect clear population differentiation and population structure within Sweden into a southern and a northern cluster. These results possibly reflect different introduction routes of barley into Sweden. We thus show that the study of historic material can be an important alternative for regions where no or little extant landrace material is available.


Next-generation sequence data demonstrate several pathogenic bee viruses in Middle East and African honey bee subspecies (Apis mellifera syriaca, Apis mellifera intermissa) as well as their cohabiting pathogenic mites (Varroa destructor).
RNA viruses are associated with honey bee (Apis mellifera) colony losses in many parts of the world. Their consequences may be exacerbated when the ectoparasite mite Varroa destructor is present in hives. While evidence of pathogenic, viral-induced disease is abundant in western honey bees (Apis mellifera mellifera) from many parts of the world, less information exists regarding the pathogen load of Apis mellifera syriaca and Apis mellifera intermissa, honey bees from the Middle East and North Africa (MENA) that play substantial roles in regional beekeeping. Here, we used next-generation sequencing to evaluate the viral populations of these subspecies and their associated mites. We found that both A. m. syriaca and A. m. intermissa, as well as the Varroa mites infecting their colonies, bear a suite of RNA viruses including major pathogenic viruses like Deformed wing virus, Acute bee paralysis virus, Black queen cell virus and Sacbrood virus, and less common viruses (e.g., bee Macula-like virus and Apis mellifera filamentous virus). The two native honey bee MENA subspecies have acquired different but overlapping suites of pathogens, which also differ, but overlap, with the suites detected in the mites. The presence of plant viruses suggests that they were acquired from foraging for pollen and nectar. Phylogenetic analysis of the above common pathogenic RNA viruses showed unexpected genetic relationships with other known strains, indicative of import to MENA from outside of the region. Our findings indicate that it is important to carefully consider the impact of the movement of queens and mobile colonies, and the effects such movement have, on the transmission of disease.


Non-Ceruloplasmin Copper Distinguishes A Distinct Subtype of Alzheimer's Disease: A Study of EEG-Derived Brain Activity.
Meta-analyses show that percentages of non-Cp-Cu-copper that is not bound to ceruloplasmin (also known as 'free' copper)-in serum are higher in Alzheimer's disease (AD) patients. Genetic heterogeneity in AD patients stratified on the basis of non-Cp-Cu cut-off sustains the existence of a copper AD metabolic subtype. Non-Cp-Cu abnormalities correlated with alterations of electroencephalographic rhythms (EEG). We aimed to determine whether an EEG-derived brain cortical rhythm's heterogeneity between two AD groups stratified on the basis of a copper marker. We assessed levels of copper, ceruloplasmin, Non-Cp-Cu, and the APOE4 genotype in 67 AD patients and compared resting EEG-derived eLORETA cortical rhythms between AD groups stratified in terms of 'Normal' and 'High' non-Cp-Cu. The High non-Cp-Cu group experienced a lower power in all bands (0.2-48 Hz) in the parietal cortices (p=0.019) and a more limited alpha band (8-13 Hz) power in the sensory lobes (temporal, occipital, and parietal p>0.05 consistently) than the Normal non-Cp-Cu AD group. When corrected for MMSE, the non-Cp-Cu levels correlated with a reduction of high-frequency brain activity (from high alpha to gamma, 10.5-48 Hz). This neurophysiological heterogeneity in EEG-derived brain cortical rhythms between the two AD groups sustains a copper AD metabolic subtype; Non-Cp-Cu is a marker of this copper AD.


Neurofilament Heavy Chain and Tau Protein Are Not Elevated in Cerebrospinal Fluid of Adult Patients with Spinal Muscular Atrophy during Loading with Nusinersen.
Nusinersen is the first approved drug for the treatment of spinal muscular atrophy (SMA). Treatment of SMA with nusinersen is based on a fixed dosing regimen. For other motoneuron diseases, such as amyotrophic lateral sclerosis (ALS), biomarkers are available for clinical diagnostics; however, no such biomarkers have yet been found for SMA. Serum and cerebrospinal fluid (CSF) samples of 11 patients with adult SMA type 3 were prospectively collected and analyzed during loading with nusinersen. Neurofilament heavy chain, tau protein, S100B protein, and neuron-specific enolase were investigated as potential biomarkers of motor neuron destruction. No significant pathological alterations in levels of neurofilament heavy chain, tau protein, or S100B protein were detected in the CSF or blood samples under baseline conditions or during loading with nusinersen. Neuron-specific enolase was marginally elevated in CSF and blood samples without significant alteration during treatment. In a mixed cohort of adult patients with SMA type 3, neurofilament heavy chain, tau protein, S100B protein, and neuron-specific enolase do not serve as potential biomarkers during the loading phase of nusinersen. The slow progression rate of SMA type 3 may not lead to detectable elevation of levels of these common markers of axonal degradation.


Postgranulomatous anetoderma associated with Takayasu's arteritis in a child.
Takayasu's arteritis (TA) is a rare chronic inflammatory arteriopathy affecting mainly the aorta and its branches. Many skin manifestations have been reported in association with this disease. Pyoderma gangrenosum and subcutaneous inflammatory lesions of the leg are the most frequent. We studied a boy with TA in whom a papular rash of the trunk preceded the onset of vascular symptoms by many years. Histologically, the lesions were superficial and consisted of middermal noncaseating tuberculoid granulomas, which progressed to atrophy and anetoderma because of elastic network disruption. Granulomas were also found in synovial tissue but not in a temporal artery biopsy specimen, which showed only intimal hyperplasia. Our observations suggest that vascular and skin lesions with elastic tissue may both result from a common granulomatous hypersensitivity process.


Contrast-dependent saturation adjustment for outdoor image enhancement.
Outdoor images captured in bad-weather conditions usually have poor intensity contrast and color saturation since the light arriving at the camera is severely scattered or attenuated. The task of improving image quality in poor conditions remains a challenge. Existing methods of image quality improvement are usually effective for a small group of images but often fail to produce satisfactory results for a broader variety of images. In this paper, we propose an image enhancement method, which makes it applicable to enhance outdoor images by using content-adaptive contrast improvement as well as contrast-dependent saturation adjustment. The main contribution of this work is twofold: (1) we propose the content-adaptive histogram equalization based on the human visual system to improve the intensity contrast; and (2) we introduce a simple yet effective prior for adjusting the color saturation depending on the intensity contrast. The proposed method is tested with different kinds of images, compared with eight state-of-the-art methods: four enhancement methods and four haze removal methods. Experimental results show the proposed method can more effectively improve the visibility and preserve the naturalness of the images, as opposed to the compared methods.


Prioritization of Pediatric Palliative Care Field-Advancement Activities in the United States: Results of a National Survey.
The field of pediatric palliative care (PPC) continues to encounter challenges and opportunities to improving access to high-quality PPC services. In early 2019, a workshop identified 11 potential "next step" actions, and subsequently a national survey-based poll of members of the PPC community was conducted to prioritize these potential actions in terms of their "actionable importance." Invitations to the survey were distributed in October 2019 to interdisciplinary PPC health care professionals via email to two major listservs, one hosted by the Section of Hospice and Palliative Medicine of the American Academy of Pediatrics, the other by the Center to Advance Palliative Care. Respondents rated the "actionable importance" of items relative to each other via a discrete choice experiment. Median importance scores are reported for each item. One hundred seventy-seven individuals responded to the survey. The majority (62.2%) were physicians, with nurses (16.4%), advanced practice nurses (7.9%), and social workers (7.3%) being the other most common responders. The top five potential actions, in descending rank order, were: Determine what parents value regarding PPC (median score of 17.8, out of a total score of all items of 100); Define and disseminate core primary PPC curriculum (median, 15.3); Develop PPC national representation strategy and tactics (median, 12.3); Create PPC-specific program development toolkit (median, 10.9); and, Analyze payment and financing ratios (median, 9.6). Those seeking to advance the field of PPC should take into account the findings from this study, which suggest that certain actions are more likely to have a beneficial impact on moving the field forward.


Imaging the water snow-line during a protostellar outburst.
A snow-line is the region of a protoplanetary disk at which a major volatile, such as water or carbon monoxide, reaches its condensation temperature. Snow-lines play a crucial role in disk evolution by promoting the rapid growth of ice-covered grains. Signatures of the carbon monoxide snow-line (at temperatures of around 20 kelvin) have recently been imaged in the disks surrounding the pre-main-sequence stars TW Hydra and HD163296 (refs 3, 10), at distances of about 30 astronomical units (au) from the star. But the water snow-line of a protoplanetary disk (at temperatures of more than 100 kelvin) has not hitherto been seen, as it generally lies very close to the star (less than 5 au away for solar-type stars). Water-ice is important because it regulates the efficiency of dust and planetesimal coagulation, and the formation of comets, ice giants and the cores of gas giants. Here we report images at 0.03-arcsec resolution (12 au) of the protoplanetary disk around V883 Ori, a protostar of 1.3 solar masses that is undergoing an outburst in luminosity arising from a temporary increase in the accretion rate. We find an intensity break corresponding to an abrupt change in the optical depth at about 42 au, where the elevated disk temperature approaches the condensation point of water, from which we conclude that the outburst has moved the water snow-line. The spectral behaviour across the snow-line confirms recent model predictions: dust fragmentation and the inhibition of grain growth at higher temperatures results in soaring grain number densities and optical depths. As most planetary systems are expected to experience outbursts caused by accretion during their formation, our results imply that highly dynamical water snow-lines must be considered when developing models of disk evolution and planet formation.


Characterization of platelet-activating factor synthesized by normal and granulocyte-macrophage colony-stimulating factor-primed human eosinophils.
Platelet-activating factor (PAF; 1-alkyl-2-acetyl-sn-glycero-3- phosphocholine) is a mediator involved in the pathogenesis of inflammatory diseases associated with tissue eosinophil infiltration. Previous studies utilizing bioassay or assaying enzymes associated with PAF biosynthesis have suggested that human eosinophils produce PAF. The present study has extended these initial studies by identifying and quantifying the different PAF molecular species and analogues synthesized by human eosinophils in response to A23187 and f-Met-Leu-Phe (FMLP). Gas chromatography-mass spectrometric analysis indicated that A23187-stimulated eosinophils produce at least three molecular species of PAF. The predominant species is 1-hexadecyl-2-acetyl-GPC (16:0) followed by 1-octadecyl-2-acetyl-GPC (18:0) and 1-octadecyl-2-acetyl-GPC (18:1). Eosinophils stimulated with FMLP produce approximately 100-fold smaller quantities of PAF relative to those produced in response to A23187 and only the 16:0 molecular species could be measured. A small percentage (comprising between 2 and 5%) of the 2-acetylated phospholipids produced by eosinophils was the 1-acyl analogue of PAF. Long-term (72 hr) incubation with granulocyte-macrophage colony-stimulating factor (GM-CSF) resulted in a three- to fourfold increase in PAF synthesis from eosinophils stimulated with FMLP, without changes in the profile of PAF molecular species or in the percentage of the 1-acyl analogue of PAF. These data indicate that human eosinophils can produce various molecular species of PAF and that this process can be quantitatively enhanced by GM-CSF.


Effects of simultaneous knockdown of HER2 and PTK6 on malignancy and tumor progression in human breast cancer cells.
Breast cancer is the most common malignancy in women of the Western world. One prominent feature of breast cancer is the co- and overexpression of HER2 and protein tyrosine kinase 6 (PTK6). According to the current clinical cancer therapy guidelines, HER2-overexpressing tumors are routinely treated with trastuzumab, a humanized monoclonal antibody targeting HER2. Approximately, 30% of HER2-overexpressing breast tumors at least initially respond to the anti-HER2 therapy, but a subgroup of these tumors develops resistance shortly after the administration of trastuzumab. A PTK6-targeted therapy does not yet exist. Here, we show for the first time that the simultaneous knockdown in vitro, compared with the single knockdown of HER2 and PTK6, in particular in the trastuzumab-resistant JIMT-1 cells, leads to a significantly decreased phosphorylation of crucial signaling proteins: mitogen-activated protein kinase 1/3 (MAPK 1/3, ERK 1/2) and p38 MAPK, and (phosphatase and tensin homologue deleted on chromosome ten) PTEN that are involved in tumorigenesis. In addition, dual knockdown strongly reduced the migration and invasion of the JIMT-1 cells. Moreover, the downregulation of HER2 and PTK6 led to an induction of p27, and the dual knockdown significantly diminished cell proliferation in JIMT-1 and T47D cells. In vivo experiments showed significantly reduced levels of tumor growth following HER2 or PTK6 knockdown. Our results indicate a novel strategy also for the treatment of trastuzumab resistance in tumors. Thus, the inhibition of these two signaling proteins may lead to a more effective control of breast cancer.


Prevalence of autism spectrum disorders - Autism and Developmental Disabilities Monitoring Network, United States, 2006.
Autism spectrum disorders (ASDs) are a group of developmental disabilities characterized by atypical development in socialization, communication, and behavior. ASDs typically are apparent before age 3 years, with associated impairments affecting multiple areas of a person's life. Because no biologic marker exists for ASDs, identification is made by professionals who evaluate a child's developmental progress to identify the presence of developmental disorders. 2006. Earlier surveillance efforts indicated that age 8 years is a reasonable index age at which to monitor peak prevalence. The identified prevalence of ASDs in U.S. children aged 8 years was estimated through a systematic retrospective review of evaluation records in multiple sites participating in the Autism and Developmental Disabilities Monitoring (ADDM) Network. Data were collected from existing records in 11 ADDM Network sites (areas of Alabama, Arizona, Colorado, Florida, Georgia, Maryland, Missouri, North Carolina, Pennsylvania, South Carolina, and Wisconsin) for 2006. To analyze changes in identified ASD prevalence, CDC compared the 2006 data with data collected from 10 sites (all sites noted above except Florida) in 2002. Children aged 8 years with a notation of an ASD or descriptions consistent with an ASD were identified through screening and abstraction of existing health and education records containing professional assessments of the child's developmental progress at health-care or education facilities. Children aged 8 years whose parent(s) or legal guardian(s) resided in the respective areas in 2006 met the case definition for an ASD if their records documented behaviors consistent with the Diagnostic and Statistical Manual of Mental Disorders, 4th edition, text revision (DSM-IV-TR) criteria for autistic disorder, pervasive developmental disorder--not otherwise specified (PDD NOS), or Asperger disorder. Presence of an identified ASD was determined through a review of data abstracted from developmental evaluation records by trained clinician reviewers. For the 2006 surveillance year, 2,757 (0.9%) of 308,038 [corrected] children aged 8 years residing in the 11 ADDM sites were identified as having an ASD, indicating an overall average prevalence of 9.0 per 1,000 population (95% confidence interval [CI] = 8.6--9.3). ASD prevalence per 1,000 children aged 8 years ranged from 4.2 in Florida to 12.1 in Arizona and Missouri, with prevalence for the majority of sites ranging between 7.6 and 10.4. For 2006, ASD prevalence was significantly lower in Florida (p<0.001) and Alabama (p<0.05) and higher in Arizona and Missouri (p<0.05) than in all other sites. The ratio of males to females ranged from 3.2:1 in Alabama to 7.6:1 in Florida. ASD prevalence varied by type of ascertainment source, with higher average prevalence in sites with access to health and education records (10.0) compared with sites with health records only (7.5). Although parental or professional concerns regarding development before age 36 months were noted in the evaluation records of the majority of children who were identified as having an ASD, the median age of earliest documented ASD diagnosis was much later (range: 41 months [Florida]-60 months [Colorado]). Of 10 sites that collected data for both the 2002 and 2006 surveillance years, nine observed an increase in ASD prevalence (range: 27%-95% increase; p<0.01), with increases among males in all sites and among females in four of 11 sites, and variation among other subgroups. In 2006, on average, approximately 1% or one child in every 110 in the 11 ADDM sites was classified as having an ASD (approximate range: 1:80-1:240 children [males: 1:70; females: 1:315]). The average prevalence of ASDs identified among children aged 8 years increased 57% in 10 sites from the 2002 to the 2006 ADDM surveillance year. Although improved ascertainment accounts for some of the prevalence increases documented in the ADDM sites, a true increase in the risk for children to develop ASD symptoms cannot be ruled out. On average, although delays in identification persisted, ASDs were being diagnosed by community professionals at earlier ages in 2006 than in 2002. These results indicate an increased prevalence of identified ASDs among U.S. children aged 8 years and underscore the need to regard ASDs as an urgent public health concern. Continued monitoring is needed to document and understand changes over time, including the multiple ascertainment and potential risk factors likely to be contributing. Research is needed to ascertain the factors that put certain persons at risk, and concerted efforts are essential to provide support for persons with ASDs, their families, and communities to improve long-term outcome.


Targeting SARS-CoV-2 Nsp12/Nsp8 interaction interface with approved and investigational drugs: an in silico structure-based approach.
In this study, the Nsp12-Nsp8 complex of SARS-CoV-2 was targeted with structure-based and computer-aided drug design approach because of its vital role in viral replication. Sequence analysis of RNA-dependent RNA polymerase (Nsp12) sequences from 30,366 different isolates were analysed for possible mutations. FDA-approved and investigational drugs were screened for interaction with both mutant and wild-type Nsp12-Nsp8 interfaces. Sequence analysis revealed that 70.42% of Nsp12 sequences showed conserved P323L mutation, located in the Nsp8 binding cleft. Compounds were screened for interface interaction, any with XP GScores lower than -7.0 kcal/mol were considered as possible interface inhibitors. RX-3117 (fluorocyclopentenyl cytosine) and Nebivolol had the highest binding affinities in both mutant and wild-type enzymes, therefore they were selected and resultant protein-ligand complexes were simulated for analysis of stability over 100 ns. Although the selected ligands had partial mobility in the binding cavity, they were not removed from the binding pocket after 100 ns. The ligand RX-3117 remained in the same position in the binding pocket of the mutant and wild-type enzyme after 100 ns MD simulation. However, the ligand Nebivolol folded and embedded in the binding pocket of mutant Nsp12 protein. Overall, FDA-approved and investigational drugs are able to bind to the Nsp12-Nsp8 interaction interface and prevent the formation of the Nsp12-Nsp8 complex. Interruption of viral replication by drugs proposed in this study should be further tested to pave the way for in vivo studies towards the treatment of COVID-19.Communicated by Ramaswamy H. Sarma.


Hepatitis C virus core protein targets 4E-BP1 expression and phosphorylation and potentiates Myc-induced liver carcinogenesis in transgenic mice.
Hepatitis C virus (HCV) is a leading cause of liver diseases including the development of hepatocellular carcinoma (HCC). Particularly, core protein has been involved in HCV-related liver pathologies. However, the impact of HCV core on signaling pathways supporting the genesis of HCC remains largely elusive. To decipher the host cell signaling pathways involved in the oncogenic potential of HCV core, a global quantitative phosphoproteomic approach was carried out. This study shed light on novel differentially phosphorylated proteins, in particular several components involved in translation. Among the eukaryotic initiation factors that govern the translational machinery, 4E-BP1 represents a master regulator of protein synthesis that is associated with the development and progression of cancers due to its ability to increase protein expression of oncogenic pathways. Enhanced levels of 4E-BP1 in non-modified and phosphorylated forms were validated in human hepatoma cells and in mouse primary hepatocytes expressing HCV core, in the livers of HCV core transgenic mice as well as in HCV-infected human primary hepatocytes. The contribution of HCV core in carcinogenesis and the status of 4E-BP1 expression and phosphorylation were studied in HCV core/Myc double transgenic mice. HCV core increased the levels of 4E-BP1 expression and phosphorylation and significantly accelerated the onset of Myc-induced tumorigenesis in these double transgenic mice. These results reveal a novel function of HCV core in liver carcinogenesis potentiation. They position 4E-BP1 as a tumor-specific target of HCV core and support the involvement of the 4E-BP1/eIF4E axis in hepatocarcinogenesis.


Anticoagulants in the prevention of venous thromboembolism.
For 50 years, the key to successfully preventing venous thrombosis (VT) or pulmonary embolism (PE) among high-risk patients has been the judicious use of anticoagulants: first through full doses of oral anticoagulants and more recently through low-dose heparin prophylaxis. Low-dose heparin has become the standard of comparison for other preventive methods, since it is relatively safe and simple, its ability to prevent approximately 65% of the subclinical VT found by leg scanning after elective general surgery is well known, and recent meta-analysis of the many pertinent published clinical trials (large and small) strongly suggests a much greater benefit: a 65% reduction in the risk of postoperative death from major PE. In addition, there are trials that have also found low-dose heparin to be effective in general medical patients, although its value in this clinical setting is much less well documented. Although several effective approaches other than low-dose heparin are available, many of these tend to be either more cumbersome (intermittent external leg compression) or probably less powerful (graded pressure elastic stockings). There are situations where low-dose heparin prophylaxis fails, most obviously after orthopaedic surgery where the use of more complex regimens, including adjusted-dose heparin treatment and various schedules of warfarin prophylaxis, becomes appropriate. Recent progress has come from the intensive clinical exploration of various low molecular weight heparin fractions or fragments which appear to be effective after once daily administration to general surgical patients and show great promise of effectiveness and safety after hip surgery. The level of warfarin effect needed for VT prophylaxis has also been reinvestigated, with trials suggesting a need for less warfarin and a lower prothrombin time effect than was previously thought to be appropriate. Given that any attempts to minimize mortality from PE in hospital patients must rely on the widespread and systematic use of simple, safe, and cost-effective preventive methods, it is hoped that these advances will help move anticoagulant prophylaxis further out of the realm of clinical research and into that of common clinical practice.


How can we strengthen pathology services in Cambodia?
Rapid economic growth and a changing disease burden have increased the demand for pathology services in Cambodia. This paper describes the status of pathological services and international support for pathology professionals in Cambodia, and discusses future needs for strengthening pathology services. In 2016, there were only four pathologists and 18 pathology technologists in Cambodia. A postgraduate course in pathology was created in 2015, and five residents became certified in 2018. Besides multinational support with lectures and practice for pathologists, the Japanese team provides on-the-job training for pathology technologists to improve slide preparation for diagnosis. A clinicopathological conference was introduced to strengthen the communication among pathologists, pathology technologists, and gynecologists. Although there is a long way to go to reach high quality pathological services, coordination among international partners needs to continue, as does the balance between human resource development for pathology professionals, to provide a higher level of care to local citizens.


Short-term memory advantage for brief durations in human APOE ε4 carriers.
The Apolipoprotein-E (APOE) ε4 gene allele, the highest known genetic risk factor for Alzheimer's disease, has paradoxically been well preserved in the human population. One possible explanation offered by evolutionary biology for survival of deleterious genes is antagonistic pleiotropy. This theory proposes that such genetic variants might confer an advantage, even earlier in life when humans are also reproductively fit. The results of some small-cohort studies have raised the possibility of such a pleiotropic effect for the ε4 allele in short-term memory (STM) but the findings have been inconsistent. Here, we tested STM performance in a large cohort of individuals (N = 1277); nine hundred and fifty-nine of which included carrier and non-carriers of the APOE ε4 gene, those at highest risk of developing Alzheimer's disease. We first confirm that this task is sensitive to subtle deterioration in memory performance across ageing. Importantly, individuals carrying the APOE ε4 gene actually exhibited a significant memory advantage across all ages, specifically for brief retention periods but crucially not for longer durations. Together, these findings present the strongest evidence to date for a gene having an antagonistic pleiotropy effect on human cognitive function across a wide age range, and hence provide an explanation for the survival of the APOE ε4 allele in the gene pool.


Differential role of cannabinoids in the pathogenesis of skin cancer.
Cannabinoids (CB) like ∆(9)-tetrahydrocannabinol (THC) can induce cancer cell apoptosis and inhibit angiogenesis. However, the use of cannabinoids for the treatment of malignant diseases is discussed controversially because of their immunomodulatory effects which can suppress anti-tumor immunity. Here we investigated the role of exogenous and endogenous cannabinoids in mouse skin cancer. First we examined the effect of THC, which binds to CB receptors (CB1, CB2), on the growth of the mouse melanoma cell lines B16 and HCmel12 in vitro and in vivo in wild type (WT) and CB1/CB2-receptor deficient mice (Cnr1/2(-/-)). Next we evaluated the role of the endogenous cannabinoid system by studying the growth of chemically induced melanomas, fibrosarcoma and papillomas in WT and Cnr1/2(-/-) mice. THC significantly inhibited tumor growth of transplanted HCmel12 melanomas in a CB receptor-dependent manner in vivo through antagonistic effects on its characteristic pro-inflammatory microenvironment. Chemically induced skin tumors developed in a similar manner in Cnr1/2(-/-) mice when compared to WT mice. Our results confirm the value of exogenous cannabinoids for the treatment of melanoma but do not support a role for the endogenous cannabinoid system in the pathogenesis of skin cancer.


Identifying adaptive genetic divergence among populations from genome scans.
The identification of signatures of natural selection in genomic surveys has become an area of intense research, stimulated by the increasing ease with which genetic markers can be typed. Loci identified as subject to selection may be functionally important, and hence (weak) candidates for involvement in disease causation. They can also be useful in determining the adaptive differentiation of populations, and exploring hypotheses about speciation. Adaptive differentiation has traditionally been identified from differences in allele frequencies among different populations, summarised by an estimate of FST. Low outliers relative to an appropriate neutral population-genetics model indicate loci subject to balancing selection, whereas high outliers suggest adaptive (directional) selection. However, the problem of identifying statistically significant departures from neutrality is complicated by confounding effects on the distribution of FST estimates, and current methods have not yet been tested in large-scale simulation experiments. Here, we simulate data from a structured population at many unlinked, diallelic loci that are predominantly neutral but with some loci subject to adaptive or balancing selection. We develop a hierarchical-Bayesian method, implemented via Markov chain Monte Carlo (MCMC), and assess its performance in distinguishing the loci simulated under selection from the neutral loci. We also compare this performance with that of a frequentist method, based on moment-based estimates of FST. We find that both methods can identify loci subject to adaptive selection when the selection coefficient is at least five times the migration rate. Neither method could reliably distinguish loci under balancing selection in our simulations, even when the selection coefficient is twenty times the migration rate.