Immunization with plant-expressed hemagglutinin protects chickens from lethal highly pathogenic avian influenza virus H5N1 challenge infection.
Highly pathogenic avian influenza (HPAI) is a striking disease in susceptible poultry, which leads to severe economic losses. Inactivated vaccines are the most widely used vaccines in avian influenza virus (AIV) vaccination programs. However, these vaccines interfere with the serological detection of wild-type AIV infections in immunized populations. The use of vaccines that allow differentiation between infected and vaccinated animals (DIVA strategy) would stop current stamping-out policies. Therefore, novel vaccination strategies are needed to allow improved protection of animals and humans against HPAI virus (HPAIV) infection. The presented study analyzed for the first time the immunogenic capacity of plant-expressed full-length hemagglutinin (rHA0) of HPAIV H5N1 in several vaccine formulations within the highly relevant host species chicken. We were able to express plant-expressed rHA0 at high levels and could show that, when administered with potent adjuvants, it is highly immunogenic and can fully protect chicken against lethal challenge infection. Real-time reverse transcription (RT)-PCR and serological tests demonstrated only marginally increased virus replication in animals vaccinated with plant-derived rHA0 compared to animals immunized with an inactivated reference vaccine. In addition, the use of plant-expressed rHA0 also allowed an easy serological differentiation of vaccinated from AIV-infected animals based on antibodies against the influenza virus NP protein.


CDK12 promotes papillary thyroid cancer progression through regulating the c-myc/β-catenin pathway.
Background: CDK12 is a potential therapeutic target in papillary thyroid cancer that regulates the c-myc/β-catenin pathway. Objective: We aimed to explore the specific mechanism of CDK12 in papillary thyroid cancer and provide a new target of cancer therapy. Methods: RT-qPCR was used to determine the CDK12 mRNA expression level. An IHC assay was performed to detect the tissue expression of CDK12. Then, we downregulated CDK12 expression in the thyroid cancer cell lines TPC-1-shCDK12 and KAT-5-shCDK12. CCK8 assays, colony formation assays, and animal xenograft models were used to evaluate the effect of CDK12 on tumorigenesis. Transwell assays and in vivo metastasis models were used to observe whether CDK12 can promote cancer metastasis. Western blotting further confirmed the mechanism of CDK12 in papillary thyroid cancer through the c-myc/β-catenin pathway. Results: Upregulated CDK12 expression in papillary thyroid cancer promoted papillary thyroid cancer carcinogenesis in vivo, and in vitro CDK12 strengthened papillary thyroid cancer (PTC) cell migration and tumor metastasis. CDK12 promoted tumor progression by regulating c-myc/β-catenin pathway activation. Conclusions: CDK12 affects the c-myc/β-catenin pathway to stimulate papillary thyroid cancer proliferation and metastasis. Inhibiting CDK12 might be a new method in papillary thyroid cancer therapy.


[Effect of different diets on quality of life in patients with metabolic syndrome].
The purpose of the research was to study influence of diets with a various ratio of carbohydrates and fatty components on modification of metabolic risk factors due to decrease of weight and abdominal adiposity, and also on quality of life of the patient. 49 males were included in the study the age 30-65 years with metabolic syndrome X. All patients had increased body mass or obesity. Hypertension of I and II stages was observed in 49.0% of cases. The estimation of results was carried out in three months after assignment of one of investigated diets. Effective reduction of body mass parameters was achieved at use of all diets within three months. Nutritional counseling was based on dietary preferences and habits of the patient to improve quality of life at observance of a diet. Common negative feature of investigated diets was occurrence of feeling of hunger that caused infringement of accuracy of observance of recommendations.


Comparison of two procedures for tracing participants in an epidemiologic cohort study.
The effectiveness and accuracy of using a commercial firm for tracing survey participants were compared with the results of an established, University of Minnesota tracing protocol. Studying a cohort of 622 survey participants ages 25-74 who were living in a single Midwestern community, we were unable to locate 142 (23%) at their original address after four years. We traced them according to the Minnesota protocol, using information obtained from contact persons (friends or relatives) and other sources. Overall, current address information was found for 96% of the participants lost to follow-up, and 113 (81%) of 139 addresses were supplied through the contact person. An independent attempt to obtain similar information using only credit bureau tracing through Associated Credit Services provided addresses for 123 of the participants. Associated Credit Services provided correct, current addresses for 39 (29%) of the 136 surviving participants (six fatal cases were excluded) and supplied correct, current addresses for a majority (58%) of the study participants found by protocol to be living in the study area and eligible for a follow-up clinic examination. We support the use of commercial services as secondary sources of address information in tracing participants after the initial use of contact persons to optimize participant contact. We recommend the use of commercial services as primary sources when contact persons are unavailable.


Hepatic resection for hepatocellular carcinoma in cirrhosis.
Hepatocellular carcinoma is one of the most common cancers worldwide. Several treatment modalities have been proposed, but hepatic resection is still considered the first-line therapeutic option for most of the patient carries of HCC. The proper selection of patients candidate to hepatic resection for HCC and the eradication whenever is possible of the intrahepatic metastases are the most crucial steps for improving the surgical outcome in HCC. This article reviews the current state of the art of the surgical treatment of HCC.


The role of Twitter in dental education: A systematic review.
The rapid growth of social media in recent years has highlighted uses beyond their original purposes, particularly in education. Twitter is a free, open access social network with high potential to enhance interactive learning. The use of Twitter in dental education has been far less investigated; therefore, the objective of this systematic review is to explore the current uses and to examine the impact of Twitter on dental education, and to analyze and predict potential models of Twitter for future application in dental training, education, and teaching. Five databases (PubMed, Embase, Medline, Scopus, and Web of Science) and the gray literature using keywords related to Twitter and dental education were searched. Articles were screened for inclusion, and two researchers independently extracted the data using a standardized data collection template and analyzed the quality of the included articles using the Medical Education Research Study Quality Instrument. Of the 121 articles identified from the initial search, 68 remained after duplications were removed. Article screening removed 61 articles leaving 7 eligible for inclusion and data extraction. Five studies were cross-sectional and two were cohort studies, and all involved survey-based designs with 998 respondents in total. Quality assessment gave a score range between 8 and 12.5 out of a total of 18 points. Our study supports the potential for Twitter as a useful learning tool in dental education. Features, including the open access nature of Twitter as well as the low level of ads and free registration, make it appealing to students as well as a useful tool for interactive learning. However, there are significant barriers to its use, including privacy and concerns about professionalism. Higher quality and greater impact research is required.


Crossing Borders Between Frontotemporal Dementia and Psychiatric Disorders: An Updated Overview.
Frontotemporal dementia (FTD) includes a group of neurocognitive syndromes, clinically characterized by altered behaviors, impairment of language proficiency, and altered executive functioning. FTD is one of the most frequently observed forms of dementia in the elderly population and the most common in presenile age. As for other subtypes of dementia, FTD incidence is constantly on the rise due to the steadily increasing age of the population, and its recognition is now becoming a determinant for clinicians. FTD and psychiatric disorders can overlap in terms of clinical presentations by sharing a common genetic predisposition and neuropathological mechanism in some cases. Nonetheless, this association is often unclear and underestimated. Since its first reports, research into FTD has constantly grown, with the identification of recent findings related to its neuropathology, genetic, clinical, and therapeutic issues. Literature is thriving on this topic, with numerous research articles published in recent years. In the present review, we aimed to provide an updated description of the clinical manifestations that link and potentially confound the diagnosis of FTD and psychiatric disorders in order to improve their differential diagnosis and early detection. In particular, we systematically reviewed the literature, considering articles specifically focused on the behavioral variant FTD, published after 2015 on the PubMed database.


Living with facial disfigurement--strategies for individuals and care management.
Individuals and families affected by craniofacial disorders have expressed dissatisfaction with their experiences in the healthcare system, with day care, and in school situations. To capture their views, focus group encounters were done in a group of young individuals with these disorders and in a group of parents whose children were affected. The aim was to synthesize their attitudes and experiences into improved strategies for parents, teenagers, and professionals in the healthcare system. Their views were compiled into a document that emphasizes the responsibilities of persons with craniofacial disorders and their parents to actively seek information on diagnosis and treatment options and to participate in decisions on therapy. The conclusion was that it is not lack of specific knowledge but rather a lack of implementation of existing recommendations that makes living with facial disfigurement difficult for many individuals and their families.


Unilateral posterior atlantoaxial transarticular screw fixation.
Bilateral posterior C 1-2 transfacet screw placement with associated posterior bone graft wiring is the accepted treatment for patients with atlantoaxial instability. This technique was modified to treat 19 patients with atlantoaxial instability and unilateral anomalies that prevented placement of a screw across the C1-2 facet. In these cases, a single contralateral transarticular screw was placed in conjunction with interspinous bone graft wiring to avoid neural or vertebral artery injury and to provide C1-2 stability. Postoperatively, all 19 patients were placed in Philadelphia collars (mean immobilization 8 weeks, range 6-12 weeks). Unilateral C1-2 facet screw fixation was needed for the following reasons: a high-riding transverse foramen of the C-2 vertebra present in 13 patients (left side in eight, right side in five), poor screw purchase in two (left side in both), screw malposition in one (left side), severe degenerative arthritis in one (right side), neurofibroma in one (right side), and fracture of the C-1 lateral mass in one (left side). Six weeks postsurgery, one patient presented with a broken screw and required occipitocervical fusion with a Steinmann pin and wire cable from the occiput to C-3 to achieve solid fusion. Solid fusions were achieved in the other 18 patients (mean follow-up period 31 months, range 14-54 months); there was no delayed screw breakage, wire breakage, or spinal instability. There were no operative or postoperative neurological or vascular complications. The authors' experience demonstrates that unilateral C1-2 facet screw fixation with interspinous bone graft wiring is an excellent alternative in the treatment of atlantoaxial instability when bilateral screw fixation is contraindicated.


Modulation of human platelet protein kinase C by endotoxic lipid A.
Lipid A is the toxic principle of lipopolysaccharide of gram-negative bacteria, which causes a spectrum of changes in blood cells and vascular cells. We now report that human platelets are directly stimulated by endotoxic lipid A that activates protein kinase C. Rapid phosphorylation of a human platelet protein of Mr 47,000, a marker of protein kinase C activation, accompanies secretion of [14C]serotonin and aggregation triggered by endotoxic lipid A. These events are time and concentration dependent, with phosphorylation reaching maximum in 2 min and the concentration of lipid A causing a 50% effect (EC50) between 12 and 15 microM. Phospholipase C activation in lipid A-stimulated platelets was not observed as judged by a lack of generation of [3H]diacylglycerol in [3H]arachidonic acid-labeled platelets and a lack of generation of [32P]-phosphatidic acid in 32PO4-labeled platelets. Lipid A did not induce formation of TXA2 as measured by radioimmunoassay for TXB2. The stimulation of human platelets and activation of protein kinase C by endotoxic lipid A was blocked by lipid X, a structural precursor of lipid A. Lipid X also blocked the stimulation of human platelets by phorbol 12-myristate 13-acetate, suggesting that lipid A, lipid X and phorbol ester share reactive site(s) on the human platelet membrane. Although lipid X inhibited thrombin-induced phosphorylation of P47 it did not suppress secretion of [14C]serotonin, indicating the role of protein kinase C-independent pathways in platelet stimulation by thrombin. The inhibitory effect of lipid X did not involve generation of cyclic AMP in human platelet membrane preparations. These results indicate that human platelets are stimulated by endotoxic lipid A, a naturally occurring biologic modifier of protein kinase C. Due to the widespread presence of this enzyme in blood cells, vascular cells, and neurons, its modulation by lipid A may represent a significant mechanism underlying hematologic and circulatory derangements observed in endotoxic shock in humans.


Physiologic oxygen enhances human embryonic stem cell clonal recovery and reduces chromosomal abnormalities.
Human embryonic stem cells (hESC) have great potential in regenerative medicine, provided that culture systems are established that maintain genomic integrity. Here we describe a comparison of the effects of culture in either physiologic oxygen (2%) or room oxygen (21%) on the hESC lines, H1, H9, and RH1. Physiologic oxygen enabled an average sixfold increase in clone recovery across the hESC lines tested (p < 0.001). FACS analysis showed that cells cultured in physiologic oxygen were significantly smaller and less granular. No significant changes had occurred in levels of SSEA4, SSEA1, TRA-1-60, or TRA-1-81. While karyotypic normalcy was maintained in both H1 and H9, the frequency of spontaneous chromosomal aberrations was significantly increased in room oxygen. This increase was not observed in physiologic oxygen. These results clearly demonstrate that physiologic oxygen culture conditions are indispensable for robust hES clone recovery and may enhance the isolation of novel hES lines and transgenic clones.


Evaluation of the patient with carcinoma of unknown origin metastatic to bone.
Metastatic carcinoma to bone of uncertain primary origin is a common clinical diagnostic scenario. I present a simple and effective staging system to identify primary malignancies. The role of clinical history, physical examination, laboratory studies, and limited radiographic studies are critical to the success of this diagnostic strategy. Needle or open biopsy frequently is needed yet usually is more effective as a confirmatory study for carcinoma or to rule out primary bone sarcomas. Patients who present with a displaced pathologic fracture require an urgent diagnostic and treatment plan that does not compromise future treatment options and provides durable skeletal reconstruction.


Phosphorylation of inositol 1,4,5-trisphosphate receptors by protein kinase B/Akt inhibits Ca2+ release and apoptosis.
Imbalance of signals that control cell survival and death results in pathologies, including cancer and neurodegeneration. Two pathways that are integral to setting the balance between cell survival and cell death are controlled by lipid-activated protein kinase B (PKB)/Akt and Ca(2+). PKB elicits its effects through the phosphorylation and inactivation of proapoptotic factors. Ca(2+) stimulates many prodeath pathways, among which is mitochondrial permeability transition. We identified Ca(2+) release through inositol 1,4,5-trisphosphate receptor (InsP(3)R) intracellular channels as a prosurvival target of PKB. We demonstrated that in response to survival signals, PKB interacts with and phosphorylates InsP(3)Rs, significantly reducing their Ca(2+) release activity. Moreover, phosphorylation of InsP(3)Rs by PKB reduced cellular sensitivity to apoptotic stimuli through a mechanism that involved diminished Ca(2+) flux from the endoplasmic reticulum to the mitochondria. In glioblastoma cells that exhibit hyperactive PKB, the same prosurvival effect of PKB on InsP(3)R was found to be responsible for the insensitivity of these cells to apoptotic stimuli. We propose that PKB-mediated abolition of InsP(3)-induced Ca(2+) release may afford tumor cells a survival advantage.


Sequencing and de novo assembly of 150 genomes from Denmark as a population reference.
Hundreds of thousands of human genomes are now being sequenced to characterize genetic variation and use this information to augment association mapping studies of complex disorders and other phenotypic traits. Genetic variation is identified mainly by mapping short reads to the reference genome or by performing local assembly. However, these approaches are biased against discovery of structural variants and variation in the more complex parts of the genome. Hence, large-scale de novo assembly is needed. Here we show that it is possible to construct excellent de novo assemblies from high-coverage sequencing with mate-pair libraries extending up to 20 kilobases. We report de novo assemblies of 150 individuals (50 trios) from the GenomeDenmark project. The quality of these assemblies is similar to those obtained using the more expensive long-read technology. We use the assemblies to identify a rich set of structural variants including many novel insertions and demonstrate how this variant catalogue enables further deciphering of known association mapping signals. We leverage the assemblies to provide 100 completely resolved major histocompatibility complex haplotypes and to resolve major parts of the Y chromosome. Our study provides a regional reference genome that we expect will improve the power of future association mapping studies and hence pave the way for precision medicine initiatives, which now are being launched in many countries including Denmark.


Strategies of adults for dealing with feelings of guilt - findings from a large web-based survey.
Feelings of guilt are common in adulthood. Depending on the situation, individuals need different strategies for dealing with these usually unpleasant experiences. The aim of this study was to provide comprehensive empirical information about such different guilt-related strategies used in adulthood. A cross-sectional web-based survey was conducted (05/2019-04/2020) collecting autobiographical information on guilt experiences. Stated strategies for dealing with feelings of guilt were classified into inductively defined categories, and the frequency of the categorized strategies was calculated (total, gender-, age-specific). Associations between applied strategies and gender and age were analyzed using Cramér's φ and V as appropriate. Analyzing statements of 579 participants (18-84 years), we differentiated 34 different categories of guilt-related strategies. The most frequently used strategies were "Self-reflection; contemplation; analyzing/reviewing things one feels guilty about" and "Repressing/ignoring/(actively) forgetting/not thinking about/bottling up the feelings of guilt". Female and male participants showed a significantly different use only in the strategy of utilizing "Professional support/therapy/counseling" (10.57% vs. 3.67%; Cramér's φ = .13, p = .001). Additionally, only a few differences were found with regard to age. Adults use a broad variety of different strategies for dealing with feelings of guilt. Strategies regulating emotional responses to the problem causing feelings of guilt seem to be at least as important as strategies targeting the problem itself. Many strategies do not serve pro-social functions. Feelings of guilt are an issue relevant for mental health care since several of the adults with feelings of guilt utilize professional help, and an even higher number may need such help.


Update on Glucose Management Among Noncritically Ill Patients Hospitalized on Medical and Surgical Wards.
Hyperglycemia is a common issue affecting inpatient care. Although this is in part because of the higher rate of hospitalization among patients with preexisting diabetes, multiple factors complicate inpatient glucose management, including acute stress from illness or surgery, erratic dietary intake, and contribution of medications. It has been repeatedly demonstrated that poorly controlled blood glucose levels are associated with negative clinical outcomes, such as increased mortality, higher rate of surgical complications, and longer length of hospital stay. Given these concerns, there has been extensive study of the optimal strategy for management of glucose levels, with the bulk of existing literature focusing on insulin therapy in the intensive care unit setting. This review shifts the focus to the general adult medical and surgical wards, using clinical guidelines and sentinel studies to describe the scientific basis behind the current basal-bolus insulin-based approach to blood sugar management among noncritically ill inpatients. Patient-centered clinical trials looking at alternative dosing regimens and insulin analog and noninsulin agents, such as glucagon-like peptide-1 agonist therapies, introduce safe and effective options in the management of inpatient hyperglycemia. Data from these studies reveal that these approaches are of comparable safety and efficacy to the traditional basal-bolus insulin regimen, and may offer additional benefit in terms of less monitoring requirements and lower rates of hypoglycemia. Although existing data are encouraging, outcome studies will be needed to better establish the clinical impact of these more recently proposed approaches in an effort to broaden and improve current clinical practices in inpatient diabetes care.


α-1,2-Mannosidase MAN1C1 Inhibits Proliferation and Invasion of Clear Cell Renal Cell Carcinoma.
Background: This study investigated the biological function of the gene MAN1C1 α-mannosidase in renal cell carcinoma. It has been reported that MAN1C1 is probably a potential tumor suppressor gene in Wilms. However, the role of MAN1C1 in human clear cell renal cell carcinoma (ccRCC) has not been reported. Methods: In this study, MAN1C1 gene over-expression was used to transfect human renal cancer cell lines 786-O and OS-RC-2 to study apoptosis and the underlying mechanisms which influence epithelial-mesenchymal transition. Results: MAN1C1 was down-regulated in ccRCC and related to the clinicopathological factors and prognosis of ccRCC. We revealed that over-expression MAN1C1 showed anti-tumor effect by inducing apoptosis, as determined by Cell Counting Kit-8 (CCK-8) assay, cell cycle analysis, and western blot analysis. What's more, MAN1C1 over-expression remarkably increased the ratio of Bax/Bcl-2 and inhibited epithelial-mesenchymal transition by increasing the expression of E-CA. In addition, the ratio of Bax/Bcl-2 and E-CA were also increased in MAN1C1 gene over-expression renal cancer cells compared with the control cells. Conclusion: We find that re-expression of silenced MAN1C1 in ccRCC cell lines inhibited cell viability, colony formation, induced apoptosis, suppressed cell invasion and migration. In conclusion, MAN1C1 is a novel functional tumor suppressor in renal carcinogenesis. This is the first time that the function of MAN1C1 gene has been verified in the renal tumor tissue so far.


Control of magnetic bearing systems via the Chebyshev polynomial-based unified model (CPBUM) neural network.
A Chebyshev polynomial-based unified model (CPBUM) neural network is introduced and applied to control a magnetic bearing systems. First, we show that the CPBUM neural network not only has the same capability of universal approximator, but also has faster learning speed than conventional feedforward/recurrent neural network. It turns out that the CPBUM neural network is more suitable in the design of controller than the conventional feedforward/recurrent neural network. Second, we propose the inverse system method, based on the CPBUM neural networks, to control a magnetic bearing system. The proposed controller has two structures; namely, off-line and on-line learning structures. We derive a new learning algorithm for each proposed structure. The experimental results show that the proposed neural network architecture provides a greater flexibility and better performance in controlling magnetic bearing systems.


Lung transplantation and survival in children with cystic fibrosis: solid statistics--flawed interpretation.
In their provocative paper, "Lung transplantation and survival in children with cystic fibrosis," Liou and colleagues state that "Prolongation of life by means of lung transplantation should not be expected in children with cystic fibrosis. A prospective, randomized trial is needed to clarify whether and when patients derive a survival and quality of life benefit from lung transplantation." Unfortunately, that conclusion is not supportable. Liou's dataset introduced bias against transplantation by using covariates obtained well before the time of transplant (when predicted survival was good) and having a cohort with lower than expected post-transplant survival than reported elsewhere. The calculated hazard ratios are based on factors that may have changed between listing and transplant, and do not reflect true benefit on a patient by patient basis. The findings of the study are contrary to other studies using similar methods. Finally, recent changes in US lung transplant allocation policy may have made the study findings moot. In contrast to Liou's suggestion to perform an ethically and logistically challenging randomized trial to verify the benefit of lung transplantation, a research agenda is recommended for pediatric lung transplantation for cystic fibrosis that focuses on developing strategies to continually reassess and maximize quality of life and survival benefit.


Heterogeneous dynamics of ionic liquids in confined films with varied film thickness.
Dynamical behavior and characteristics of 1-butyl-3-methylimidazolium hexafluorophosphate ([BMIM][PF6]) ionic liquid (IL) in confined films with varied film thickness have been investigated using atomistic molecular dynamics simulations. Simulation results indicate that the dynamics of confined ionic groups in interfacial regions is highly heterogeneous and depends strongly on their relative layered positions in the confined IL film. The dynamics and relaxation times of the ionic groups in the bulk region of confined IL films are very similar to that in the corresponding pure bulk simulation. In contrast, the diffusion of the corresponding ionic groups slows down and can be characterized by slaved diffusion while the corresponding relaxation times increase remarkably as these ionic groups come closer to the neutral graphene surface. While at the IL-vacuum interface, the diffusion and relaxation of terminal carbon atoms of butyl chains of [BMIM] cations are much faster than in other layers of the confined IL film and in the bulk region of the simulation system without confinement, due to their librated motion in this interfacial region. The dynamical heterogeneity of the confined ionic groups is intrinsically related to microscopic ionic structures and the orientational preference of [BMIM][PF6] ion pairs in interfacial regions.


Effect of norepinephrine on gluconeogenesis in perfused livers of cold-exposed rats.
The effects of norepinephrine and glucagon on gluconeogenesis were studied in hemoglobin-free perfused liver from rats kept for 1-20 days at 4 degrees C. When rats were starved for 24 h at 4 degrees C, the plasma glucose level of rats exposed to cold for 5, 10, and 20 days was significantly higher than that of rats for 1 day, but hepatic glycogen decreased to the same level in all groups. In the isolated perfused liver, basal rates of oxygen consumption and glucose production increased slightly through 5 days of cold exposure and returned to control levels after 20 days of cold exposure. The rates of glucose production from lactate, pyruvate, sorbitol, and glycerol increased by 20-30% after 5 days of cold exposure. The stimulation of gluconeogenesis from these substrates by norepinephrine and phenylephrine increased markedly at all time periods from 1 to 20 days in the cold, with a maximum at 5 days. The stimulation of glycogenolysis by norepinephrine was not affected by cold exposure. The response to catecholamines decreased markedly in liver perfused with calcium-free medium and/or with phentolamine. The stimulation of gluconeogenesis by glucagon increased only in rats exposed to cold for 20 days. The results obtained suggest that the stimulation of hepatic gluconeogenesis by cold is due to an alpha-adrenergic response, and the activation occurs beyond the interaction of norepinephrine with its receptor.


Nitric oxide, malaria, and anemia: inverse relationship between nitric oxide production and hemoglobin concentration in asymptomatic, malaria-exposed children.
The cause of the anemia associated with chronic, intermittent, asymptomatic, low-level parasitemia in children in malaria-endemic endemic areas is not well understood. Nitric oxide (NO) decreases erythropoiesis, and it is likely an important mediator of anemia of chronic disease. Production of NO is decreased in acute uncomplicated and cerebral malaria, but it is increased in asymptomatic Tanzanian children (with or without parasitemia). We hypothesized that chronic overproduction of NO in these asymptomatic children contributes to the anemia associated with subclinical/subpatent malaria. In 44 fasting, asymptomatic, malaria-exposed, Tanzanian children, NO production (measured using fasting urine NOx excretion) was inversely associated with hemoglobin concentration (P = 0.03, controlling for age and gender). Using multiple linear regression, hemoglobin concentration was negatively associated with parasitemia (P = 0.005). After controlling for age and parasitemia, NO was no longer an independent predictor of anemia. One of the mechanisms of parasite-related anemia in such children may be through the adverse hematologic effects of parasite-induced NO production.


Individual and contextual factors associated with low childhood immunisation coverage in sub-Saharan Africa: a multilevel analysis.
In 2010, more than six million children in sub-Saharan Africa did not receive the full series of three doses of the diphtheria-tetanus-pertussis vaccine by one year of age. An evidence-based approach to addressing this burden of un-immunised children requires accurate knowledge of the underlying factors. We therefore developed and tested a model of childhood immunisation that includes individual, community and country-level characteristics. We conducted multilevel logistic regression analysis of Demographic and Health Survey data for 27,094 children aged 12-23 months, nested within 8,546 communities from 24 countries in sub-Saharan Africa. According to the intra-country and intra-community correlation coefficient implied by the estimated intercept component variance, 21% and 32% of the variance in unimmunised children were attributable to country- and community-level factors respectively. Children born to mothers (OR 1.35, 95%CI 1.18 to 1.53) and fathers (OR 1.13, 95%CI 1.12 to 1.40) with no formal education were more likely to be unimmunised than those born to parents with secondary or higher education. Children from the poorest households were 36% more likely to be unimmunised than counterparts from the richest households. Maternal access to media significantly reduced the odds of children being unimmunised (OR 0.94, 95%CI 0.94 to 0.99). Mothers with health seeking behaviours were less likely to have unimmunised children (OR 0.56, 95%CI 0.54 to 0.58). However, children from urban areas (OR 1.12, 95% CI 1.01 to 1.23), communities with high illiteracy rates (OR 1.13, 95% CI 1.05 to 1.23), and countries with high fertility rates (OR 4.43, 95% CI 1.04 to 18.92) were more likely to be unimmunised. We found that individual and contextual factors were associated with childhood immunisation, suggesting that public health programmes designed to improve coverage of childhood immunisation should address people, and the communities and societies in which they live.


Waste-Based Pigments for Application in Ceramic Glazes and Stoneware Bodies.
The use of wastes, some of them hazards, as raw materials of ceramic pigments has been a way to diminish their environmental impact, to economically valorize them, and to face the depletion of virgin raw materials. In this work were prepared pigments having in their composition only industrial wastes: Cr/Ni electroplating (ES), and sludges from the cutting of natural stones-marble (MS) and granite (GS). The prepared mixtures were calcined at three temperatures (1100, 1200, and 1300 °C) and the obtained powders were characterized by XRD and UV-vis. Their coloring strength and thermal stability were assessed by adding them to different ceramic substrates: glazes (transparent bright and opaque matte) and a stoneware paste. The CIEL*a*b* coordinates of the fired materials were measured. The developed pigments are thermally stable and exhibit good tinting power, originating nicely colored and defect-free ceramic materials.


A region of the myosin rod important for interaction with paramyosin in Caenorhabditis elegans striated muscle.
The precise arrangement of molecules within the thick filament, as well as the mechanisms by which this arrangement is specified, remains unclear. In this article, we have exploited a unique genetic interaction between one isoform of myosin heavy chain (MHC) and paramyosin in Caenorhabditis elegans to probe the molecular interaction between MHC and paramyosin in vivo. Using chimeric myosin constructs, we have defined a 322-residue region of the MHC A rod critical for suppression of the structural and motility defects associated with the unc-15(e73) allele. Chimeric constructs lacking this region of MHC A either fail to suppress, or act as dominant enhancers of, the e73 phenotype. Although the 322-residue region is required for suppression activity, our data suggest that sequences along the length of the rod also play a role in the isoform-specific interaction between MHC A and paramyosin. Our genetic and cell biological analyses of construct behavior suggest that the 322-residue region of MHC A is important for thick filament stability. We present a model in which this region mediates an avid interaction between MHC A and paramyosin in parallel arrangement in formation of the filament arms.


Healing of excisional wound in alloxan induced diabetic sheep: A planimetric and histopathologic study.
Healing of skin wound is a multi-factorial and complex process. Proper treatment of diabetic wounds is still a major clinical challenge. Although diabetes mellitus can occur in ruminants, healing of wounds in diabetic ruminants has not yet been investigated. The aim of this study was to evaluate healing of ovine excisional diabetic wound model. Eight 4-month-old Iranian Makoui wethers were equally divided to diabetic and nondiabetic groups. Alloxan monohydrate (60 mg kg(-1), IV) was used for diabetes induction. In each wether, an excisional wound was created on the dorsum of the animal. Photographs were taken in distinct times for planimetric evaluation. Wound samples were taken on day 21 post-wounding for histopathologic evaluations of epidermal thickness, number of fibroblasts and number of new blood vessels. The planimetric study showed slightly delay in wound closure of diabetic animals, however, it was not significantly different from nondiabetic wounds (p ≥ 0.05). Furthermore, epidermal thickness, number of fibroblasts and number of blood vessels were significantly lower in diabetic group (p < 0.05). We concluded that healing of excisional diabetic wounds in sheep may be compromised, as seen in other species. However, contraction rate of these wounds may not be delayed due to metabolic features of ruminants and these animals might go under surgeries without any serious concern. However, healing quality of these wounds may be lower than normal wounds.


Guanylate cyclase in Dictyostelium discoideum with the topology of mammalian adenylate cyclase.
The core of adenylate and guanylate cyclases is formed by an intramolecular or intermolecular dimer of two cyclase domains arranged in an antiparallel fashion. Metazoan membrane-bound adenylate cyclases are composed of 12 transmembrane spanning regions, and two cyclase domains which function as a heterodimer and are activated by G-proteins. In contrast, membrane-bound guanylate cyclases have only one transmembrane spanning region and one cyclase domain, and are activated by extracellular ligands to form a homodimer. In the cellular slime mould, Dictyostelium discoideum, membrane-bound guanylate cyclase activity is induced after cAMP stimulation; a G-protein-coupled cAMP receptor and G-proteins are essential for this activation. We have cloned a Dictyostelium gene, DdGCA, encoding a protein with 12 transmembrane spanning regions and two cyclase domains. Sequence alignment demonstrates that the two cyclase domains are transposed, relative to these domains in adenylate cyclases. DdGCA expressed in Dictyostelium exhibits high guanylate cyclase activity and no detectable adenylate cyclase activity. Deletion of the gene indicates that DdGCA is not essential for chemotaxis or osmo-regulation. The knock-out strain still exhibits substantial guanylate cyclase activity, demonstrating that Dictyostelium contains at least one other guanylate cyclase.


Impact of input data uncertainty on environmental exposure assessment models: A case study for electromagnetic field modelling from mobile phone base stations.
With the increased availability of spatial data and computing power, spatial prediction approaches have become a standard tool for exposure assessment in environmental epidemiology. However, such models are largely dependent on accurate input data. Uncertainties in the input data can therefore have a large effect on model predictions, but are rarely quantified. With Monte Carlo simulation we assessed the effect of input uncertainty on the prediction of radio-frequency electromagnetic fields (RF-EMF) from mobile phone base stations at 252 receptor sites in Amsterdam, The Netherlands. The impact on ranking and classification was determined by computing the Spearman correlations and weighted Cohen's Kappas (based on tertiles of the RF-EMF exposure distribution) between modelled values and RF-EMF measurements performed at the receptor sites. The uncertainty in modelled RF-EMF levels was large with a median coefficient of variation of 1.5. Uncertainty in receptor site height, building damping and building height contributed most to model output uncertainty. For exposure ranking and classification, the heights of buildings and receptor sites were the most important sources of uncertainty, followed by building damping, antenna- and site location. Uncertainty in antenna power, tilt, height and direction had a smaller impact on model performance. We quantified the effect of input data uncertainty on the prediction accuracy of an RF-EMF environmental exposure model, thereby identifying the most important sources of uncertainty and estimating the total uncertainty stemming from potential errors in the input data. This approach can be used to optimize the model and better interpret model output.


Ethical problems in pediatrics: what does the setting of care and education show us?
Pediatrics ethics education should enhance medical students' skills to deal with ethical problems that may arise in the different settings of care. This study aimed to analyze the ethical problems experienced by physicians who have medical education and pediatric care responsibilities, and if those problems are associated to their workplace, medical specialty and area of clinical practice. A self-applied semi-structured questionnaire was answered by 88 physicians with teaching and pediatric care responsibilities. Content analysis was performed to analyze the qualitative data. Poisson regression was used to explore the association of the categories of ethical problems reported with workplace and professional specialty and activity. 210 ethical problems were reported, grouped into five areas: physician-patient relationship, end-of-life care, health professional conducts, socioeconomic issues and health policies, and pediatric teaching. Doctors who worked in hospitals as well as general and subspecialist pediatricians reported fewer ethical problems related to socioeconomic issues and health policies than those who worked in Basic Health Units and who were family doctors. Some ethical problems are specific to certain settings: those related to end-of-life care are more frequent in the hospital settings and those associated with socioeconomic issues and public health policies are more frequent in Basic Health Units. Other problems are present in all the setting of pediatric care and learning and include ethical problems related to physician-patient relationship, health professional conducts and the pediatric education process. These findings should be taken into consideration when planning the teaching of ethics in pediatrics. This research article didn't reports the results of a controlled health care intervention. The study project was approved by the Institutional Ethical Review Committee (Report CEP-HIJG 032/2008).


Assessing Family Planning Service Quality And User Experiences In Social Franchising Programme - Case Studies From Two Rural Districts In Pakistan.
Studies have documented the impact of quality family planning services on improved contraceptive uptake and continuation, however, relatively little is known about their quality of service provision especially in the context of social franchising. This study examined the quality of clinical services and user experiences among two models in franchised service providers in rural Pakistan. This facility-based assessment was carried out during May-June 2015 at the 20 randomly selected social franchise providers from Chakwal and Faisalabad. In our case, a franchise health facility was a private clinic (mostly) run by a single provider, supported by an assistant. Within the selected health facilities, a total 39 user-provider interactions were observed and same users were interviewed separately. Most of the health facilities were in the private sector. Comparatively, service providers at Greenstar Social Marketing/Population Services International (GSM/PSI) model franchised facilities had higher number of rooms and staff employed, with more providers' ownership. Quality of service indices showed high scores for both Marie Stopes Society (MSS) and GSM/PSI franchised providers. MSS franchised providers demonstrated comparative edge in terms of clinical governance, better method mix and they were more user-focused, while PSI providers offered broader range of non-FP services. Quality of counselling services were similar among both models. Service providers performed well on all indicators of interpersonal care however overall low scores were noted in technical care. For both models, service providers attained an average score of 6.7 (out of the maximum value of 8) on waste disposal mechanism, supplies 12.5 (out of the maximum value of 15), user-centred facility 2.7 (out of the maximum value of 4), and clinical governance 6.5 (out of the maximum value of 11) and respecting clients' privacy. The exit interviews yielded high user satisfaction in both service models. The findings seem suggesting that the MSS and GSM/PSI service providers were maintaining high quality standards in provision of family planning information, services, and commodities but overall there was not much difference between the two models in terms of quality and satisfaction. The results demonstrate that service quality and client satisfaction are an important determinant of use of clinical contraceptive methods in Pakistan.


A putative glutathione-binding site in CdZn-metallothionein identified by equilibrium binding and molecular-modelling studies.
Glutathione (GSH) has been found to form a complex with both vertebrate and invertebrate copper-metallothionein (CuMT) [Freedman, Ciriolo and Peisach (1989) J. Biol. Chem. 264, 5598-5605; Brouwer and Brouwer-Hoexum (1991) Arch. Biochem. Biophys. 290, 207-213]. In this paper we report on the interaction of GSH with CdZnMT-I and CdZnMT-II from rabbit liver and with CdMT-I from Blue crab hepatopancreas. Ultrafiltration experiments showed that all three MTs combined with GSH. The measured binding data for the three MTs could be described by a single binding isotherm. The GSH/MT stoichiometry was 1.4 +/- 0.3 and Kdiss. = 14 +/- 6 microM. Partially Zn-depleted MT does not significantly bind GSH, indicating that the GSH-binding site is located on MT's Zn-containing N-terminal domain. The putative GSH-binding site on rabbit liver MT was investigated using molecular-graphics analysis. A cleft on the MT's N-terminal domain, which has the labile Zn-2 at its base, could easily accommodate GSH. Cysteine-ligand exchange between the terminal (non-bridging) Cys-26, bound to Zn-2, and the cysteine in GSH is stereochemically possible. Based on these considerations a model of MT-GSH was built in which GSH's cysteine replaces Cys-26 as a terminal Zn-2 ligand. This complex was energy-minimized by molecular-mechanics calculations, taking into account computed partial electrostatic charges on all atoms, including Cd and Zn. These calculations showed that the MT-GSH complex was thermodynamically more stable than MT, due to favourable non-bonded, electrostatic and van der Waals interactions. Six hydrogen bonds can form between GSH and MT. The average pairwise root-mean-square deviations (RMSD) of the metals in energy-minimized MT and MT-GSH, compared with the metals in the crystal structure, were 0.0087 +/- 0.0028 nm (0.087 +/- 0.028 A) and 0.0168 +/- 0.0087 nm (0.168 +/- 0.087 A) respectively. The RMSD values for the polypeptide-backbone alpha carbons were 0.0136 +/- 0.0060 nm (0.136 +/- 0.060 A) and 0.0491 +/- 0.0380 nm (0.491 +/- 0.380 A) respectively. No other docking sites for GSH were found. The energy-minimized structure of an MT-2-mercaptoethanol complex was somewhat less stable than the native MT domain, attesting to the specificity of the MT-GSH interaction. The possible physiological significance of the MT-GSH interaction is discussed.


A matched-cohort analysis of 192 pancreatic anaplastic carcinomas and 960 pancreatic adenocarcinomas: A 13-year North American experience using the National Cancer Data Base (NCDB).
Anaplastic pancreatic carcinoma (APC) is a rare and poorly characterized disease. We sought to compare the clinical characteristics and outcomes of APC to pancreatic adenocarcinoma (PDAC). The American National Cancer Data Base was queried for patients with resected APC and PDAC using histologic and operative codes. APC cases were matched 1:5 with PDACs based on age, sex, pathologic tumor stage, operative margin status, lymph node positivity ratio, and use of adjuvant chemotherapy. After 1:5 matching, 192 APCs and 960 PDACs were analyzed. When comparing APC vs PDAC the median tumor size was 45 mm (interquartile range, 33-60) vs 30 mm (interquartile range, 23-40; P < .001), and metastatic nodal disease was present in 40.6% and 38.0% of the cases (P = .25), respectively. APC cases were distributed equally between the head and the body/tail region of the pancreas (50%), while PDAC cases were located mainly in the head of the pancreas (75%; P < .001). Although the resected APC group had a lesser survival during the first year after the diagnosis (51% vs 69%; P = .029), the overall survival was similar in the 2 groups, with 21.6% vs 17.4% alive at 5 years, respectively for APC and PDAC (P = .32). Subgroup analysis of patients with APC with (n = 18) versus those without (n = 80) osteoclastlike giant cells showed a greater 5-year survival (50% versus 15%, P < .001). Patients with resected APC tend to present with large tumors equally distributed between the head and body/tail of the pancreas. While APC is thought to have a more aggressive biology, our matched analysis showed similar overall survival compared with PDAC. The presence of osteoclastlike giant cells portends a significantly better prognosis compared with other histologic features of APCs.


Determination of crude protein in animal feed, forage, grain, and oilseeds by using block digestion with a copper catalyst and steam distillation into boric acid: collaborative study.
A collaborative study was conducted to evaluate the repeatability and reproducibility of an extension of AOAC Official Method 991.20, Nitrogen (Crude) in Milk, to animal feed, forage (plant tissue), grain, and oilseed materials. Test portions are digested in an aluminum block at 420 degrees C in sulfuric acid with potassium sulfate and a copper catalyst. Digests are cooled and diluted, and concentrated sodium hydroxide is added to neutralize the acid and make the digest basic; the liberated ammonia is distilled by using steam distillation. The liberated ammonia is trapped in a weak boric acid solution and titrated with a stronger standardized acid, hydrochloric acid; colorimetric endpoint detection is used. Fourteen blind samples were sent to 13 collaborators in the United States, Denmark, Sweden, Germany, and the United Kingdom. Recoveries of nitrogen from lysine, tryptophan, and acetanilide were 86.8, 98.8, and 100.1%, respectively. The within-laboratory relative standard deviation (RSDr, repeatability) ranged from 0.40 to 2.38% for crude protein. The among-laboratories (including within-) relative standard deviation (RSD(R), reproducibility) ranged from 0.44 to 2.38%. It is recommended that the method be adopted First Action by AOAC INTERNATIONAL. A lower concentration (1% H3BO3) of trapping solution was compared with the concentration specified in the original protocol (4% H3BO3) and was found comparable for use in an automatic titration system in which titration begins automatically as soon as distillation starts. The Study Directors recommend that 1% H3BO3 as an optional alternative to 4% boric acid trapping solution be allowed for automatic titrators that titrate throughout the distillation.


The complexity hypothesis revisited: connectivity rather than function constitutes a barrier to horizontal gene transfer.
Horizontal gene transfer (HGT) is a prevalent and a highly important phenomenon in microbial species evolution. One of the important challenges in HGT research is to better understand the factors that determine the tendency of genes to be successfully transferred and retained in evolution (i.e., transferability). It was previously observed that transferability of genes depends on the cellular process in which they are involved where genes involved in transcription or translation are less likely to be transferred than metabolic genes. It was further shown that gene connectivity in the protein-protein interaction network affects HGT. These two factors were shown to be correlated, and their influence on HGT is collectively termed the "Complexity Hypothesis". In this study, we used a stochastic mapping method utilizing advanced likelihood-based evolutionary models to quantify gene family acquisition events by HGT. We applied our methodology to an extensive across-species genome-wide dataset that enabled us to estimate the overall extent of transfer events in evolution and to study the trends and barriers to gene transferability. Focusing on the biological function and the connectivity of genes, we obtained novel insights regarding the "complexity hypothesis." Specifically, we aimed to disentangle the relationships between protein connectivity, cellular function, and transferability and to quantify the relative contribution of each of these factors in determining transferability. We show that the biological function of a gene family is an insignificant factor in the determination of transferability when proteins with similar levels of connectivity are compared. In contrast, we found that connectivity is an important and a statistically significant factor in determining transferability when proteins with a similar function are compared.


Pretreatment cognitive deficits and treatment effects on attention in childhood absence epilepsy.
To determine the neurocognitive deficits associated with newly diagnosed untreated childhood absence epilepsy (CAE), develop a model describing the factorial structure of items measuring academic achievement and 3 neuropsychological constructs, and determine short-term differential neuropsychological effects on attention among ethosuximide, valproic acid, and lamotrigine. Subjects with newly diagnosed CAE entering a double-blind, randomized controlled clinical trial had neuropsychological testing including assessments of general intellectual functioning, attention, memory, executive function, and achievement. Attention was reassessed at the week 16-20 visit. At study entry, 36% of the cohort exhibited attention deficits despite otherwise intact neurocognitive functioning. Structural equation modeling of baseline neuropsychological data revealed a direct sequential effect among attention, memory, executive function, and academic achievement. At the week 16-20 visit, attention deficits persisted even if seizure freedom was attained. More subjects receiving valproic acid (49%) had attention deficits than subjects receiving ethosuximide (32%) or lamotrigine (24%) (p = 0.0006). Parental assessment did not reliably detect attention deficits before or after treatment (p < 0.0001). Children with CAE have a high rate of pretreatment attentional deficits that persist despite seizure freedom. Rates are disproportionately higher for valproic acid treatment compared with ethosuximide or lamotrigine. Parents do not recognize these attentional deficits. These deficits present a threat to academic achievement. Vigilant cognitive and behavioral assessment of these children is warranted. This study provides Class I evidence that valproic acid is associated with more significant attentional dysfunction than ethosuximide or lamotrigine in children with newly diagnosed CAE.


Intensity modulation methods for proton radiotherapy.
The characteristic Bragg peak of protons or heavy ions provides a good localization of dose in three dimensions. Through their ability to deliver laterally and distally shaped homogenous fields, protons have been shown to be a precise and practical method for delivering highly conformal radiotherapy. However, in an analogous manner to intensity modulation for photons, protons can be used to construct dose distributions through the application of many individually inhomogeneous fields, but with the localization of dose in the Bragg peak providing the possibility of modulating intensity within each field in two or three dimensions. We describe four different methods of intensity modulation for protons and describe how these have been implemented in an existing proton planning system. As a preliminary evaluation of the efficacy of these methods, each has been applied to an example case using a variety of field combinations. Dose-volume histogram analysis of the resulting dose distributions shows that when large numbers of fields are used, all techniques exhibit both good target homogeneity and sparing of neighbouring critical structures, with little difference between the four techniques being discerned. As the number of fields is decreased, however, only a full 3D modulation of individual Bragg peaks can preserve both target coverage and sparing of normal tissues. We conclude that the 3D method provides the greatest flexibility for constructing conformal doses in challenging situations, but that when large numbers of beam ports are available, little advantage may be gained from the additional modulation of intensity in depth.


The Effects of Virtual Reality Nonphysical Mental Training on Coordination and Skill Transfer in Healthy Adults.
Mental training is a promising method to improve motor skills. However, transfer of these improvements to different skills or functional activities is still unclear. The purpose of this study was to investigate the effects of mental balance training programs on motor coordination and skill transfer. Randomized controlled trial. Fifty-seven healthy adults (28 females and 29 males) aged between 18 and 25 years participated in this study. Participants were randomly assigned to 3 groups: virtual reality (VR) mental training group, conventional mental training group, and control group. The training program included action observation and motor imagery practice with balance exercise videos. The VR mental training group trained with a VR head-mounted display and the conventional mental training group trained with a nonimmersive computer monitor for 30 minutes, 3 days per week, for 4 weeks. Coordination skills were tested with 2 separate custom-made obstacle course tests (OCT-1 and OCT-2). OCT tests included crouching, turning, leaning, stepping over, changing direction, walking on various surfaces, or using repeated hand and arm movement tasks. OCT-1 was used to investigate the effects of mental exercises on coordination skills, and OCT-2 to investigate transfer effects for novel tasks. Test time (total and corrected) and error types (minor, major, and total) were recorded. Touching an obstacle without changing its position was classified as a minor error, and changing its position was a major error. OCT-1 test time and number of errors significantly decreased in the VR mental training and conventional mental training groups, but not in the control group. The number of minor errors was only decreased in the VR mental training group. For OCT-2, total and corrected time were not significantly different between the groups. However, both training groups were significantly superior to the control group for all types of errors. Our findings suggest that both training interventions can significantly improve coordination and skill transfer test results. In addition, VR mental training may have some advantages over conventional mental training. These findings are promising for the use of mental training for prevention and rehabilitation in special populations.


Characteristics of patients prescribed angiotensin-converting enzyme inhibitors, angiotensin receptor blockers, or the combination at an urban medical center.
The realization that angiotensin-converting enzyme (ACE) inhibitors do not provide complete blockade of angiotensin II synthesis has resulted in an increased use of combinations of ACE inhibitors and angiotensin receptor blockers (ARBs). This study examines the characteristics of patients in whom this combination was prescribed. Seventy-two patients diagnosed with primary hypertension and receiving an ACE inhibitor, an ARB, or their combinations were included. A retrospective review using outcome variables of mean arterial pressure (MAP), and changes between pretreatment MAP and post-treatment MAP were compared between groups. Statistical analysis was performed with SPSS statistical software. Analysis of variance (ANOVA) with Tukey's post hoc analysis was performed on continuous variables. Chi-square analysis was performed on categorical variables. Multivariate linear regression was performed to determine the best predictors of post-treatment MAP. There were no significant differences between the groups in pre- or post-treatment MAP. Patients on combination therapy with an ACE and ARB agent tended to be on more antihypertensive medications and tended to be diabetic. All treatment groups had similar blood pressure control and changes in MAP regardless of treatment. These findings suggest that combination ARB and ACE inhibitor therapy is a strategy being used for diabetics with difficult-to-control hypertension, although we cannot determine from our study whether this is primarily for blood pressure control or for renal protection. Whether combining an ACE inhibitor and ARB for blood pressure control alone is supported by the literature may be debatable. Further studies should evaluate the efficacy of such intervention to control hypertension. 1. Antihypertensive therapies using ACE inhibitors with ARBs are gaining popularity. 2. This retrospective chart review was completed to examine the characteristics of patients on monotherapy and of patients on combination therapy with ACE inhibitors and ARB agents. 3. This study suggests a tendency toward combined ARB and ACE inhibitor therapy in patients with diabetes who are on multiple antihypertensive medications.


Glucose Overload Inhibits Glutamatergic Synaptic Transmission: A Novel Role for CREB-Mediated Regulation of Synaptotagmins 2 and 4.
Glucose metabolism derangement is critically involved in the age-related memory loss but the underlying molecular mechanisms are still poorly understood. In a mouse model of type 1 diabetes we found memory impairment associated with inhibition of the transcription factor CREB and alteration of pre- and post-synaptic protein expression in the hippocampus. Accordingly, glucose excess negatively affected activity-dependent CREB phosphorylation and CREB-mediated mRNA expression of synaptic proteins in hippocampal primary neurons. Specifically, glucose excess inhibited the activity-dependent recruitment of CREB on the regulatory sequences of synaptotagmin (SYT) 2 and 4 promoters and the expression of SYT4 protein. As a result, high glucose affected both the frequency of miniature excitatory postsynaptic currents and NMDA receptor-mediated currents in autaptic hippocampal neuronal cultures. Collectively, our findings highlight novel mechanisms underlying hyperglycaemia-related memory loss, including CREB-dependent downregulation of synaptotagmin expression.


Adipose browning response to burn trauma is impaired with aging.
BACKGROUNDThe incidence of burn injuries in older patients is dramatically increasing as the population of older people grows. Despite the increased demand for elderly burn care, the mechanisms that mediate increased morbidity and mortality in older trauma patients are unknown. We recently showed that a burn injury invokes white adipose tissue browning that leads to a substantially increased hypermetabolic response associated with poor outcomes. Therefore, the aim of this study was to determine the effect of age on the metabolic adipose response of browning after a burn injury.METHODOne hundred and seventy patients with burn injury admitted to the Ross Tilley Burn Centre were prospectively enrolled and grouped by age as older (≥50 years) and young (≤35 years). Adipose tissue and sera were collected and analyzed for browning markers and metabolic state via histology, gene expression, and resting energy expenditure assays.RESULTSWe found that older patients with burn injury lacked the adipose browning response, as they showed significant reductions in uncoupling protein 1 (UCP1) expression. This failure of the browning response was associated with reduced whole-body metabolism and decreased survival in older patients with burn injury. Mechanistically, we found that the adipose of both aged patients after burn trauma and aged mice after a burn showed impairments in macrophage infiltration and IL-6, key immunological regulators of the browning process after a severe trauma.CONCLUSIONTargeting pathways that activate the browning response represents a potential therapeutic approach to improve outcomes after burn trauma for elderly patients.FUNDINGNIH (R01-GM087285-01), Canadian Institutes of Health Research (grant no. 123336), and Canada Foundation for Innovation Leaders Opportunity Fund (no. 25407).


Evaluation of a new surgical approach for the treatment of clomiphene citrate-resistant infertility in polycystic ovary syndrome: laparoscopic ovarian multi-needle intervention.
Laparoscopic ovarian drilling (LOD) is the accepted second-line treatment for clomiphene citrate-resistant anovulatory infertility in polycystic ovary syndrome (PCOS). Although multiple pregnancy rates are reduced with ovarian drilling procedures, postoperative adhesion formation is a potential complication in up to 85% of the women subjected to laparoscopic destructive ovarian procedures. Our objective was to determine the effectiveness of a new, specially designed laparoscopic device and technique that might enable treatment for patients with anovulatory PCOS with less trauma and fewer postoperative adhesions. Thirty-five infertile clomiphene citrate-resistant women with PCOS were included. Seventeen women underwent laparoscopic ovarian multi-needle intervention (LOMNI), and 18 women received step-up ovulation induction treatment with recombinant follicle-stimulating hormone followed by intrauterine insemination for three cycles. Patients were followed for a period of 6 months after either laparoscopic surgery or the initiation of ovulation induction therapy. Outcome measures were cycle regularity, pregnancy rate, safety, postoperative adhesion formation, and cost effectiveness. There were no significant differences between the two groups in terms of age, body-mass index, duration of infertility, and basal cycle-day 2 hormone levels. Significant improvement in cycle regularity (p <.01) was found after LOMNI. Cumulative pregnancy rates (35.3% in the LOMNI group vs 33.3% in the ovulation induction group) did not differ between the groups. No adverse events following surgery were noted. Moderate ovarian hyperstimulation syndrome and multiple pregnancies occurred in four and two patients, respectively, in the ovulation induction group. Eight nonpregnant women in the LOMNI group underwent repeat laparoscopy at the end of the follow-up period. No adhesion formation attributable to LOMNI was observed in any of those eight women. The cost of LOMNI was significantly (p <.001) lower than the ovulation induction treatment. In conclusion, LOMNI may be a safe, inexpensive, and effective procedure for the treatment of CC-resistant infertility in patients with PCOS. It seems to preserve the beneficial effects and probably omits unwanted effects (such as adhesion formation) of LOD.


National survey on school-based fluoride mouth-rinsing programme in Japan: regional spread conditions from preschool to junior high school in 2010.
We surveyed the state of implementation of the school-based fluoride mouth-rinsing programme (S-FMR) in schools in Japan from March 2010. Questionnaires on the implementation status of S-FMR in each type of school (including preschool and kindergarten) were sent by post to the oral health administration departments of all 47 prefectures and 89 cities (18 ordinance-designated cities, 23 special wards, 41 core cities and seven public health centres in ordinance-designated cities) with public health centres. The S-FMR implementation rate was low, at only 11% of all schools in Japan and only 6% of all participating school children aged 4-14 years. In many regions, the S-FMR was implemented more widely and received higher participation from children in either elementary schools and junior high schools or preschools and kindergartens. Inter-prefectural disparities were seen in S-FMR implementation, as some prefectures and cities did not include topical fluoride application in their health promotion plans, and some local public bodies did not include targets for fluoride mouth-rinsing. To reduce this disparity in Japan where systemic fluoride application is not performed, each local public body must consider implementing the S-FMR as a public health measure. We propose using the results of this survey as basic data for formulating S-FMR goals (numerical targets) and adopting S-FMR as a concrete measure in the second Healthy Japan 21, to be launched in the fiscal year for 2013, and within the basic matters of the Act Concerning the Promotion of Dental and Oral Health.


The effects from DNA extraction methods on the evaluation of microbial diversity associated with human colonic tissue.
Potentially valuable sources of DNA have been extracted from human colonic tissues and are retained in biobanks throughout the world, and might be re-examined to better understand host-microbe interactions in health and disease. However, the published protocols for DNA extraction typically used by gastroenterologists have not been systematically compared in terms of their recovery of the microbial fraction associated with colonic tissue. For this reason, we examined how three different tissue DNA extraction methods (the QIAGEN AllPrep DNA/RNA kit, salting out and high molecular weight (HMW) methods of DNA extraction) employed in past clinical trials, and the repeated bead beating and column (RBB+C) method might impact the recovery of microbial DNA from colonic tissue, using a custom designed phylogenetic microarray for gut bacteria and archaea. All four methods produced very similar profiles of the microbial diversity, but there were some differences in probe signal intensities, with the HMW method producing stronger probe intensities for a subset of the Firmicutes probes including Clostridium and Streptococcus spp. Real-time PCR analysis revealed that the HMW and RBB+C extracted DNA contained significantly more DNA of Firmicutes origin and that the different DNA extraction methods also gave variable results in terms of host DNA recovery. All of the methods tested recovered DNA from the archaeal community although there were some differences in probe signal intensity. Based on these findings, we conclude that while all four methods are efficacious at releasing microbial DNA from biopsy tissue samples, the HMW and RBB+C methods of DNA extraction may release more DNA from some of the Firmicutes bacteria associated with colonic tissue. Thus, DNA archived in biobanks could be suitable for retrospective profiling analyses, provided the caveats with respect to the DNA extraction method(s) used are taken into account.


Immunologic changes in Ostertagia ostertagi-infected calves treated strategically with an anthelmintic.
Immunomodulation by Ostertagia ostertagi and the effects of strategic anthelmintic treatments were studied in 3 groups of 6 calves each: an infected group, inoculated with 100,000 third-stage larvae (L3)/calf at 1 and 28 days; an infected-treated group, similarly inoculated, but treated with ivermectin 9 days after each inoculation; and a noninfected control group. Calves were grazed on 2 previously clean pastures, with the infected and infected-treated calves sharing 1 pasture. All calves were inoculated at 12 weeks with Brucella abortus and infectious bovine rhinotracheitis vaccines, and at 13 weeks with a soluble O ostertagi L3 extract (SLE). Infected and infected-treated calves had lower blood lymphocyte responses to phytohemagglutinin than did control calves at 2, 4, and 14 weeks, whereas the infected calves had higher responses at 6 and 8 weeks (P < 0.05). Responses to pokeweed mitogen followed a similar pattern, whereas those to SLE were low in all calves. Infected and infected-treated calves had increased values of IgG1 to excretory-secretory antigens of L4 larvae, but no changes in the values of IgG1 to SLE. They also had lower values of IgG and IgM to B abortus and antibodies to infectious bovine rhinotracheitis at 13 weeks. The infected-treated calves, however, had higher blood lymphocyte and monocyte counts and lower eosinophil count than did infected calves. They also had reduced abomasal mucosa thickness, abomasal wall weight, and abomasal lymph node weight than infected calves. It was concluded that O ostertagi causes nonspecific suppression of cellular and humoral immunity, hypersensitivity reactions coexisted with immunosuppression, and strategic anthelmintic treatments attenuated hypersensitivity reactions and initiated better immune responses.


Sex is never the same: men's perspectives on refusing circumcision from an in-depth qualitative study in Kwaluseni, Swaziland.
Faced with an HIV prevalence of 31% among 18- to 49-year-olds, Swaziland developed a male circumcision policy in 2009, following compelling scientific evidence from three randomised controlled trials. Utilising United States Agency for International Development funds, the state set out to circumcise 80% of adult men in 2011. Only 8667 of the targeted 150,000 men were circumcised during the campaign. This paper presents findings from a 2012 to 2013 in-depth qualitative study among Swazi men. Methods included 13 focus group discussions, 20 in-depth interviews, 16 informal interviews and participant observation. We argue that the campaign's failure can be partly explained by the fact that circumcision was perceived as a threat to Swazi masculinities, a factor hardly considered in the planning of the intervention. Results show that men believed circumcision resulted in reduced penis sensitivity, reduced sexual pleasure and adverse events such as possible mistakes during surgery and post-operative complications that could have negative effects on their sexual lives. Given the conflicting state of scientific data about the effects of circumcision on sexuality or sexual pleasure, this study addresses important lacunae, while also demonstrating the need for more research into the relationship between sexuality, masculinity and health interventions seeking to involve men.


Identification of a novel heterozygous PTH1R variant in a Chinese family with incomplete penetrance.
Mutations in PTH1R are associated with Jansen-type metaphyseal chondrodysplasia (JMC), Blomstrand osteochondrodysplasia (BOCD), Eiken syndrome, enchondroma, and primary failure of tooth eruption (PFE). Inheritance of the PTH1R gene can be either autosomal dominant or autosomal recessive, indicating the complexity of the gene. Our objective was to identify the phenotypic differences in members of a family with a novel PTH1R mutation. The proband was a 13-year, 6-month-old girl presenting with short stature, abnormal tooth eruption, skeletal dysplasia, and midface hypoplasia. The brother and father of the proband presented with short stature and abnormal tooth eruption. High-throughput sequencing was performed on the proband, and the variant was confirmed in the proband and other family members by Sanger sequencing. Amino acid sequence alignment was performed using ClustalX software. Three-dimensional structures were analyzed and displayed using the I-TASSER website and PyMOL software. High-throughput genome sequencing and Sanger sequencing validation showed that the proband, her father, and her brother all carried the PTH1R (NM_000316) c.1393G>A (p.E465K) mutation. The c.1393G>A (p.E465K) mutation was novel, as it has not been reported in the literature database. According to the American College of Medical Genetics and Genomics (ACMG) guidelines, the p.E465K variant was considered to have uncertain significance. Biological information analysis demonstrated that this identified variant was highly conserved and highly likely pathogenic. We identified a novel heterozygous mutation in the PTH1R gene leading to clinical manifestations with incomplete penetrance that expands the spectrum of known PTH1R mutations.


A Cross-Sectional Study on the Effect of Chronic Noise Exposure on the Vestibular Function of Traffic Policemen and Automobile Drivers.
Noise is a preventable occupational hazard for certain professions like automobile drivers and traffic police personnel. The harmful auditory effects of noise are well known. However, little is known about the status of the vestibular function in chronic noise exposure without noise induced hearing loss. Our objective was to assess the vestibular function in chronic noise exposure. : The study was conducted with a sample size of 242 (chronic noise exposure group - 121, group without chronic noise exposure - 121). Noise estimation was carried out across various traffic intersections to assess the noise exposure levels of the exposed group. All participants underwent a detailed vestibular evaluation in the clinical vestibulometry laboratory. There was no difference in nystagmus, saccades, caloric function between the two groups. The latency and amplitude of vestibular evoked myogenic potentials (VEMP) were similar in both the groups. However, dynamic posturography showed a significant difference in the Adaptation test between the two groups (P < 0.05). We also found a statistically significant difference between the static and dynamic subjective visual vertical (SVV) and the dynamic visual acuity (DVA) between the two groups (P < 0.05). We did not find any clinical evidence of vestibular dysfunction in the noise exposed group. However, the statistical significance of SVV and DVA as seen in this study needs to be evaluated further as an early marker for vestibular dysfunction. It remains to be seen whether the statistically significant prolongation is reversible after the noise exposure is withdrawn.


Diet-induced shifts in the gut microbiota influence anastomotic healing in a murine model of colonic surgery.
Host diet and gut microbiota interact to contribute to perioperative complications, including anastomotic leak (AL). Using a murine surgical model of colonic anastomosis, we investigated how diet and fecal microbial transplantation (FMT) impacted the intestinal microbiota and if a predictive signature for AL could be determined. We hypothesized that a Western diet (WD) would impact gut microbial composition and that the resulting dysbiosis would correlate with increased rates of AL, while FMT from healthy, lean diet (LD) donors would reduce the risk of AL. Furthermore, we predicted that surgical outcomes would allow for the development of a microbial preclinical translational tool to identify AL. Here, we show that AL is associated with a dysbiotic microbial community characterized by increased levels of Bacteroides and Akkermansia. We identified several key taxa that were associated with leak formation, and developed an index based on the ratio of bacteria associated with the absence and presence of leak. We also highlight a modifiable connection between diet, microbiota, and anastomotic healing, potentially paving the way for perioperative modulation by microbiota-targeted therapeutics to reduce AL.


Continuity of care for patients with chronic disease: a registry-based observational study from Norway.
Continuity of care (CoC) is accepted as a core value of primary care and is especially appreciated by patients with chronic conditions. Nevertheless, there are few studies investigating CoC for these patients across levels of healthcare. This study aims to investigate CoC for patients with somatic chronic diseases, both with regular general practitioners (RGPs) and across care levels. We conducted a registry-based observational study by using nationwide consultation data from Norwegian general practices, out-of-hours services, hospital outpatient care, and private specialists with public contracts. Patients with diabetes mellitus (type I or II), asthma, chronic obstructive pulmonary disease, or heart failure in 2012, who had ≥2 consultations with these diagnoses during 2014 were included. CoC was measured during 2014 by using the usual provider of care (UPC) index and Bice-Boxerman continuity of care score (COCI). Both indices have a value between 0 and 1. Patients with diabetes mellitus comprised the largest study population (N = 79,165) and heart failure the smallest (N = 4,122). The highest mean UPC and COCI were measured for patients with heart failure, 0.75 and 0.77, respectively. UPC increased gradually with age for all diagnoses, while COCI showed this trend only for asthma. Both indices had higher values in urban areas. Our findings suggest that CoC in Norwegian healthcare system is achieved for a majority of patients with chronic diseases. Patients with heart failure had the highest continuity with their RGP. Higher CoC was associated with older age and living in urban areas.


CD44 Contributes to the Regulation of MDR1 Protein and Doxorubicin Chemoresistance in Osteosarcoma.
Osteosarcoma is the most common type of pediatric bone tumor. Despite great advances in chemotherapy during the past decades, the survival rates of osteosarcoma patients remain unsatisfactory. Drug resistance is one of the main reasons, leading to treatment failure and poor prognosis. Previous reports correlated expression of cluster of differentiation 44 (CD44) with drug resistance and poor survival of osteosarcoma patients, however the underlying mechanisms are poorly defined. Here, we investigated the role of CD44 in the regulation of drug chemoresistance, using osteosarcoma cells isolated from mice carrying a mutation of the tumor suppressor neurofibromatosis type 2 (Nf2) gene. CD44 expression was knocked-down in the cells using CRISPR/Cas9 approach. Subsequently, CD44 isoforms and mutants were re-introduced to investigate CD44-dependent processes. Sensitivity to doxorubicin was analyzed in the osteosarcoma cells with modified CD44 expression by immunoblot, colony formation- and WST-1 assay. To dissect the molecular alterations induced by deletion of Cd44, RNA sequencing was performed on Cd44-positive and Cd44-negative primary osteosarcoma tissues isolated from Nf2-mutant mice. Subsequently, expression of candidate genes was evaluated by quantitative reverse transcription PCR (qRT-PCR). Our results indicate that CD44 increases the resistance of osteosarcoma cells to doxorubicin by up-regulating the levels of multidrug resistance (MDR) 1 protein expression, and suggest the role of proteolytically released CD44 intracellular domain, and hyaluronan interactions in this process. Moreover, high throughput sequencing analysis identified differential regulation of several apoptosis-related genes in Cd44-positive and -negative primary osteosarcomas, including p53 apoptosis effector related to PMP-22 (Perp). Deletion of Cd44 in osteosarcoma cells led to doxorubicin-dependent p53 activation and a profound increase in Perp mRNA expression. Overall, our results suggest that CD44 might be an important regulator of drug resistance and suggest that targeting CD44 can sensitize osteosarcoma to standard chemotherapy.


Reduction of dietary obesity in aP2-Ucp transgenic mice: physiology and adipose tissue distribution.
We seek to determine whether increased energy dissipation in adipose tissue can prevent obesity. Transgenic mice with C57BL6/J background and the adipocyte lipid-binding protein (aP2) gene promoter directing expression of the mitochondrial uncoupling protein (UCP) gene in white and brown fat were used. Physiologically, UCP is essential for nonshivering thermogenesis in brown fat. Mice were assigned to a chow or a high-fat (HF) diet at 3 mo of age. Over the next 25 wk, gains of body weight were similar in corresponding subgroups (n = 6-8) of female and male mice: 4-5 g in chow nontransgenic and transgenic, 20 g in HF nontransgenic, and 9-11 g in HF transgenic mice. The lower body weight gain in the HF transgenic vs. nontransgenic mice corresponded to a twofold lower feed efficiency. Gonadal fat was enlarged, but subcutaneous white fat was decreased in the transgenic vs. nontransgenic mice in both dietary conditions. The results suggest that UCP synthesized from the aP2 gene promoter is capable of reducing dietary obesity.


The site of the molecular defect in the thyroid gland of the hyt/hyt mouse: abnormalities in the TSH receptor-G protein complex.
The hyt/hyt mouse has a severe and pervasive primary inherited hypothyroidism with significantly depressed serum T4, elevated serum and pituitary TSH, and reduced thyroid gland iodide uptake. Previous ultrastructural and histologic analysis of the hyt/hyt thyroid gland along with these biochemical abnormalities support an inherited defect in TSH responsiveness of the hyt/hyt thyroid gland. In order to evaluate the potential site of the defect in the hyt/hyt mouse, we have studied the hyt/hyt gland and hyt/hyt TSH from a biochemical and molecular standpoint. Based on demonstrated bioactivity of hyt/hyt serum in the McKenzie bioassay, this reduced responsiveness to TSH in the hyt/hyt mouse is not due to reduced bioactivity of hyt/hyt TSH or a major structural abnormality in the hyt/hyt TSH molecule. In comparison to hyt/ + euthyroid littermates and +/+ BALB/cBY progenitor strain mice, the hyt/hyt mouse demonstrates a twofold reduction in thyroid gland basal cAMP and a markedly diminished response of adenylyl cyclase to exogenous TSH. However, hyt/hyt cAMP production is equivalent to the euthyroid mice after stimulation of thyroid glands by forskolin, cholera toxin, PGE1, and isoproterenol. These results support a defect in the TSH-G protein-adenylyl cyclase system in the hyt/hyt thyroid gland. Specifically, these findings suggest that the hyt/hyt mouse has a defect in TSH responsivity due to an inherited defect in the thyroid gland TSH receptor molecule. Since the hyt/hyt gland makes T3 and T4 but at diminished levels, the proposed defect in the TSH receptor would still impart partial function. Both hyt/hyt and euthyroid hyt/ + littermates make TSH receptor mRNAs of 5500 and 2400 base pairs. This suggests that the receptor defect does not represent a major structural abnormality of the gene. The receptor defect could represent a reduction in receptor number, receptor-TSH affinity, or TSH receptor-G protein coupling. The specificity of this effect on adenylyl cyclase-cAMP is shown by the reduction of TSH-cAMP regulated thyroid peroxidase (TPO) and thyroglobulin mRNAs in the hyt/hyt thyroid gland. Given the importance of TPO and thyroglobulin in normal thyroid hormone synthesis, the reductions in TPO and thyroglobulin mRNAs in the hyt/hyt thyroid gland may underlie the significant decrease in thyroid hormone production by the hyt/hyt mouse.


Using Storybooks to Teach Children About Illness Transmission and Promote Adaptive Health Behavior - A Pilot Study.
Although there is a large and growing literature on children's developing concepts of illness transmission, little is known about how children develop contagion knowledge before formal schooling begins and how these informal learning experiences can impact children's health behaviors. Here, we asked two important questions: first, do children's informal learning experiences, such as their experiences reading storybooks, regularly contain causal information about illness transmission; and second, what is the impact of this type of experience on children's developing knowledge and behavior? In Study 1, we examined whether children's commercial books about illness regularly contain contagion-relevant causal information. In Study 2, we ran a pilot study examining whether providing children with causal information about illness transmission in a storybook can influence their knowledge and subsequent behavior when presented with a contaminated object. The results from Study 1 suggest that very few (15%) children's books about illness feature biological causal mechanisms for illness transmission. However, results from Study 2 suggest that storybooks containing contagion-relevant explanations about illness transmission may encourage learning and avoidance of contaminated objects. Altogether, these results provide preliminary data suggesting that future research should focus on engaging children in learning about contagion and encouraging adaptive health behaviors.


Molecular basis for elliptocytosis associated with glycophorin C and D deficiency in the Leach phenotype.
Glycophorin C (GPC) and glycophorin D (GPD) are highly glycosylated integral membrane proteins of human erythrocytes encoded by the same gene and associated with expression of Gerbich blood group system antigens. GPC/D deficiency (the Leach phenotype) is a rare condition usually found after identification of Gerbich blood group system antibodies in persons undergoing prenatal or pretransfusion evaluation. In all cases, the Leach phenotype has been associated with elliptocytosis. Characterization of the molecular basis of this phenotype in three previously uninvestigated families has shown that the most common genetic basis of GPC/D deficiency is deletion of exons 3 and 4 of the GPC gene. However, in one family, the Leach phenotype appeared due to a deletion of one nucleotide in exon 3, causing a frameshift mutation in the messenger RNA and premature generation of a stop codon. The GPC and GPD protein sequences are therefore interrupted in the extracellular domain and probably intracellularly degraded.


Acrolein modification impairs key functional features of rat apolipoprotein E: identification of modified sites by mass spectrometry.
Apolipoprotein E (apoE), an antiatherogenic apolipoprotein, plays a significant role in the metabolism of lipoproteins. It lowers plasma lipid levels by acting as a ligand for the low-density lipoprotein receptor (LDLr) family of proteins, in addition to playing a role in promoting macrophage cholesterol efflux in atherosclerotic lesions. The objective of this study is to examine the effect of acrolein modification on the structure and function of rat apoE and to determine the sites and nature of modification by mass spectrometry. Acrolein is a highly reactive aldehyde, which is generated endogenously as one of the products of lipid peroxidation and is present in the environment in pollutants such as tobacco smoke and heated oils. In initial studies, acrolein-modified apoE was identified by immunoprecipitation using an acrolein-lysine specific antibody in the plasma of 10-week old male rats that were exposed to filtered air (FA) or low doses of environmental tobacco smoke (ETS). While both groups displayed acrolein-modified apoE in the lipoprotein fraction, the ETS group had higher levels in the lipid-free fraction compared with the FA group. This observation provided the rationale to further investigate the effect of acrolein modification on rat apoE at a molecular level. Treatment of recombinant rat apoE with a 10-fold molar excess of acrolein resulted in (i) a significant decrease in lipid-binding and cholesterol efflux abilities, (ii) impairment in the LDLr- and heparin-binding capabilities, and (iii) significant alterations in the overall stability of the protein. The disruption in the functional abilities is attributed directly or indirectly to acrolein modification yielding an aldimine adduct at K149 and K155 (+38); a propanal adduct at K135 and K138 (+56); an N(ε)-(3-methylpyridinium)lysine (MP-lysine) at K64, K67, and K254 (+76), and an N(ε)-(3-formyl-3,4-dehydropiperidino)lysine (FDP-lysine) derivative at position K68 (+94), as determined by matrix-assisted laser desorption/ionization-time of flight/time of flight mass spectrometry (MALDI-TOF/TOF MS). The loss of function may also be attributed to alterations in the overall fold of the protein as noted by changes in the guanidine HCl-induced unfolding pattern and to protein cross-linking. Overall, disruption of the structural and functional integrity of apoE by oxidative modification of essential lysine residues by acrolein is expected to affect its role in maintaining plasma cholesterol homeostasis and lead to dysregulation in lipid metabolism.


Newspaper and wire service coverage of tobacco farmers.
This study examined print media coverage of tobacco farmers from the perspective of agenda setting, or the extent to which information is available to the public and perceived as important. A content analysis of 743 articles published between January 1, 1995 and June 30, 1997 was completed. The number of articles increased from 1995 to 1997. Of the topics analyzed, articles on tobacco settlement (7.1% of total) and diversification (15.6% of total) were the least prevalent. Because the settlement discussions did not occur until 1997 (when it comprised 26.4% of the total in the first 6 months), diversification was consistently the least covered topic. The two most frequent topics covered were tobacco companies (36.2%) and the tobacco price support program (32.3%). Except for one 6 month interval, there were substantially more articles in local/regional publications than in national publications. Public health professionals have called for tobacco farmers to diversify to non-tobacco enterprises. Yet, there is little discussion of diversification in print media. Without more attention to diversification, the public and policy makers will be ill-informed about opportunities and obstacles in this regard.


Policy Responses to the Addiction Crisis.
The COVID-19 pandemic is just one of two public health crises the new Biden administration will confront. The addiction crisis is the other. The opioid epidemic has already killed more Americans than World Wars I and II combined. And it is but the most visible sign of a broader population health challenge that includes methamphetamine, cocaine, benzodiazepines, and alcohol. This article presents practical legislative and executive actions that are required for addressing these challenges. The authors focus on two broad policy challenges: (1) improving financing and delivery of treatment for substance use disorders, and (2) reducing population exposure to addictive and lethal substances. Through both of these channels, a portfolio of well-implemented, evidence-informed policies can save many thousands of lives every year.


Comparison of intracranial self-administration of ethanol within the posterior ventral tegmental area between alcohol-preferring and Wistar rats.
A previous study indicated that selectively bred alcohol-preferring (P) rats self-administered ethanol (EtOH) directly into the ventral tegmental area (VTA), whereas the alcohol-nonpreferring line did not. Wistar rats will also self-administer EtOH directly into the posterior VTA. Because Wistar rats also have a low preference for EtOH solutions but self-inject EtOH into the VTA, this study was undertaken to test the hypothesis that there is an association between EtOH preference and sensitivity of the VTA to the reinforcing effects of EtOH. Adult P and Wistar rats were assigned to groups that received one of the following concentrations of EtOH: 0, 50, 75, 100, 150, or 200 mg/100 ml. Rats were connected to the microinjection system, placed into two-lever (active and inactive) experimental chambers, and given EtOH for the first four sessions (acquisition), artificial cerebrospinal fluid for sessions 5 and 6 (extinction), and EtOH again in session 7 (reinstatement). Responding on the active lever produced a 100-nl injection of the infusate. P rats self-infused 75 to 200 mg/100 ml EtOH and demonstrated lever discrimination, whereas Wistar rats reliably self-infused only 150 and 200 mg/100 ml EtOH. Both P and Wistar rats reduced responding on the active lever when artificial cerebrospinal fluid (aCSF) was substituted for EtOH and reinstated responding in session 7 when EtOH was restored, although P rats demonstrated a very robust enhancement of responding for 100 and 150 mg/100 ml EtOH, and this was not found for Wistar rats. These results suggest that, compared with Wistar rats, the posterior VTA of P rats was more sensitive to the reinforcing effects of EtOH. Furthermore, the reinstatement data suggest that the posterior VTA of P rats underwent neuronal alterations as a result of prior EtOH exposure and extinction that changed the reinforcing effects of EtOH within this region.


Affordable Care Act and healthcare delivery: A comparison of California and Florida hospitals and emergency departments.
The Affordable Care Act (ACA) has expanded access to health insurance for millions of Americans, but the impact of Medicaid expansion on healthcare delivery and utilization remains uncertain. To determine the early impact of the Medicaid expansion component of ACA on hospital and ED utilization in California, a state that implemented the Medicaid expansion component of ACA and Florida, a state that did not. Analyze all ED encounters and hospitalizations in California and Florida from 2009 to 2014 and evaluate trends by payer and diagnostic category. Data were collected from State Inpatient Databases, State Emergency Department Databases and the California Office of Statewide Health Planning and Development. Hospital and ED encounters. Population-based study of California and Florida state residents. Implementation of Medicaid expansion component of ACA in California in 2014. Changes in ED visits and hospitalizations by payer, percentage of patients hospitalized after an ED encounter, top diagnostic categories for ED and hospital encounters. In California, Medicaid ED visits increased 33% after Medicaid expansion implementation and self-pay visits decreased by 25% compared with a 5.7% increase in the rate of Medicaid patient ED visits and a 5.1% decrease in rate of self-pay patient visits in Florida. In addition, California experienced a 15.4% increase in Medicaid inpatient stays and a 25% decrease in self pay stays. Trends in the percentage of patients admitted to the hospital from the ED were notable; a 5.4% decrease in hospital admissions originating from the ED in California, and a 2.1% decrease in Florida from 2013 to 2014. We observed a significant shift in payer for ED visits and hospitalizations after Medicaid expansion in California without a significant change in top diagnoses or overall rate of these ED visits and hospitalizations. There appears to be a shift in reimbursement burden from patients and hospitals to the government without a dramatic shift in patterns of ED or hospital utilization.