Meaning and agency in discussing end-of-life care: a study of elderly veterans' values and interpretations.
The authors of this exploratory study used textual analysis of transcribed interviews to examine the mental constructs that individuals form around advance care terminology and to learn how elderly veterans conceptualize the language used in the Veterans Administration advance directive. They found that respondents often negotiated meaning by drawing on rigid schemas, specific mental constructs already in place: The Lord's Will, Machine Talk, Being a Burden, and Being Productive. The authors also examined the transcripts for agency. In addition to assigning external agency for end-of-life care decisions, respondents often expressed a complex interaction of "self" and "other" agency. These results challenge us to develop communication methods that allow patients to claim agency and participate fully in decisions regarding their health care, especially at the end of life.


Cell Microarray Technologies for High-Throughput Cell-Based Biosensors.
Due to the recent demand for high-throughput cellular assays, a lot of efforts have been made on miniaturization of cell-based biosensors by preparing cell microarrays. Various microfabrication technologies have been used to generate cell microarrays, where cells of different phenotypes are immobilized either on a flat substrate (positional array) or on particles (solution or suspension array) to achieve multiplexed and high-throughput cell-based biosensing. After introducing the fabrication methods for preparation of the positional and suspension cell microarrays, this review discusses the applications of the cell microarray including toxicology, drug discovery and detection of toxic agents.


Seizures Following Cranioplasty: Risk Factors and Prevention Exploration.
The aim of this study was to identify risk factors and explore the possible prevention measures for seizures following cranioplasty. The authors performed a retrospective review of 142 consecutive patients who underwent cranioplasty following craniectomy for trauma or cerebral hemorrhage in Dezhou People's Hospital between January 2010 and January 2017. Patients who were diagnosed with aneurysms, arteriovenous malformations, cerebral infarction, and tumors (14), had seizures prior to cranioplasty (9) and those lost to follow-up (7) were excluded. Patients did not use antiepilepsy drugs from treatment of postcranioplasty seizures. The median follow-up time was 51.11 ± 31.59 months (range: 17-98 months). Analyses were performed on a database tracking age, sex, reason for craniectomy, operative time, time between operations, presence of dural substitute, diabetic status, hypertensive status, tobacco use, alcohol use, location of cranioplasty, classification, time and times of seizures following cranioplasty development. One hundred twelve patients met study criteria. The overall rate of seizures following cranioplasty was 35.7% (40 out of 112 patients). There were no statistically significant associations between postcranioplasty seizures and sex, age, location of cranioplasty, cranioplasty materials, or dural substitute used in craniectomy. Postcranioplasty seizure frequency differed significantly according to reasons for depressive craniectomy. The incidence of postcranioplasty seizures was significantly higher in the first year than in later years. Incidence decreased progressively in subsequent years. Incidence of seizures following cranioplasty was associated with the reason for depressive craniectomy.


A perspective view of sodium iodide symporter research and its clinical implications.
The sodium iodide symporter (NIS) is an intrinsic plasma membrane protein that mediates active iodide transport into the thyroid gland and into several extrathyroidal tissues, in particular the lactating mammary gland. Cloning and molecular characterization of the NIS have allowed the investigation of its key role in thyroid physiology as well as its potential pathophysiological and therapeutic implications in benign and malignant thyroid diseases. Similarly, elucidating the mechanisms underlying the regulation of NIS in lactating mammary gland and breast cancer, in which more than 80% of cases express endogenous NIS, may lead to findings that have novel implications for pathophysiology and therapy. Two approaches may, in the future, pave the way to extend the use of radioiodide treatment to nonthyroidal cancer. One is based on the reinduction of endogenous NIS expression in thyroid and breast cancer by targeting the main mechanisms involving tumoral transformation and dedifferentiation. The other is based on the application of NIS as a novel cytoreductive gene therapy strategy. NIS offers the unique advantage that it can be used both as a reporter and as a therapeutic gene, so that it is possible to image, monitor, and treat the tumor with radioiodide, just as in differentiated thyroid cancer. This review summarizes the main recent findings in NIS research that have a direct impact on diagnosis and therapeutic management.


Long-term outcomes following sirolimus conversion after renal transplantation.
Long-term outcomes following renal transplantation remain limited due to chronic progressive injury partly as a result of calcineurin inhibitor (CNI) toxicity. Thus, patients have been converted to non-CNI immunosuppressives despite the lack of evidence of long-term benefits from CNI free therapy. We now report our 10-year experience converting patients with well functioning transplants from CNI to sirolimus. We retrospectively analyzed outcomes of patients receiving continuous CNI based therapy (CNI, n = 309) or who were switched to sirolimus within the first year of post-transplantation (CONV, n = 54). The groups were similar for most recipient, graft and donor characteristics, however, diabetes was more common in the CNI group and statin use was more frequent in the CONV group. The average time to conversion was 7.2 months and the creatinine level at the time of switching was 1.4 mg/dl. Ten year graft and patient survival rates were equivalent in both groups. There were no differences in the causes of death or graft loss in both groups. Renal function was available for 5 years posttransplant and was no different between groups. Thus, there is no evidence that routinely switching patients with well functioning renal allografts to sirolimus from CNI based immunosuppression provides long-term benefit.


Enhanced intestinal absorption of asenapine maleate by fabricating solid lipid nanoparticles using TPGS: elucidation of transport mechanism, permeability across Caco-2 cell line and in vivo pharmacokinetic studies.
The aim of the present investigation was to fabricate and evaluate solid lipid nanoparticles (SLNs) of asenapine maleate (AM) to improve its oral bioavailability (BA). AM-SLNs were prepared by high speed homogenization followed by ultrasonication technique. The resultant SLNs exhibited particle size, zeta potential and entrapment efficiency of 114.3 ± 3.5 nm, -12.9 ± 3.8 mV, and 84.10% ± 2.90% respectively. In vitro release study of AM-SLNs showed 9.23% ± 2.72% and 92.09% ± 3.40% release of AM in pH 1.2 medium and phosphate buffer pH 6.8, respectively, indicating higher potential of lymphatic uptake. Cell viability study using Caco-2 cell line indicated non-toxicity of the carriers and drug. The uptake of AM-SLNs across Caco-2 cell line was time and energy dependent exhibiting clathrin-claveole mediated endocytosis transport. Cellular uptake of Coumarin loaded SLNs was effectively increased as compared to the dye solution. The pharmacokinetic results in rats showed 50.19-fold improvement in BA of AM after fabrication of SLNs. Collectively, all these findings demonstrated effectiveness of SLNs to improve therapeutic efficacy of AM in the treatment of schizophrenia.


An efficient strategy for TALEN-mediated genome engineering in Drosophila.
In reverse genetics, a gene's function is elucidated through targeted modifications in the coding region or associated DNA cis-regulatory elements. To this purpose, recently developed customizable transcription activator-like effector nucleases (TALENs) have proven an invaluable tool, allowing introduction of double-strand breaks at predetermined sites in the genome. Here we describe a practical and efficient method for the targeted genome engineering in Drosophila. We demonstrate TALEN-mediated targeted gene integration and efficient identification of mutant flies using a traceable marker phenotype. Furthermore, we developed an easy TALEN assembly (easyT) method relying on simultaneous reactions of DNA Bae I digestion and ligation, enabling construction of complete TALENs from a monomer unit library in a single day. Taken together, our strategy with easyT and TALEN-plasmid microinjection simplifies mutant generation and enables isolation of desired mutant fly lines in the F1 generation.


[Analysis of particle size characteristics of road sediments in Beijing Olympic Park].
Particle size analysis of road sediment collected in October and November in Beijing Olympic Park indicates that most of the sediments are 76-830 μm; the grain size of the sediments in the area of large population flow is mainly coarse but the grain size in the area of large traffic volume is fine relatively while most of the sediments are <300 p.m. Moreover, sediments of size range <300 μm can be easily accumulated on the road with moderate traffic density. The results demonstrate that the effect of pedestrian flow on the composition of the particles is unobvious and the main influences are the traffic density, extensive construction. With the length of dry period increasing, the content of sediments of size range >300 μm decreases and the content of sediments of size range < 150 μm increases, however, the change of the content of sediments of size range 150-300 μm is not obvious. The results indicate that the effectiveness of the road sediment removal depends on the length of dry period, and the accumulation of different size particles varies differently under the different dry days. Compared with the stone road, surface particles can accumulate on the asphalt road more easily as the accumulation of particles is affected by the road material significantly. Therefore, to reduce the urban surface water pollution, it is necessary to improve the design of park road such as using the stone road, which can decrease the roughness of the road.


Allelic variation of the Tas1r3 taste receptor gene selectively affects taste responses to sweeteners: evidence from 129.B6-Tas1r3 congenic mice.
The Tas1r3 gene encodes the T1R3 receptor protein, which is involved in sweet taste transduction. To characterize ligand specificity of the T1R3 receptor and the genetic architecture of sweet taste responsiveness, we analyzed taste responses of 129.B6-Tas1r3 congenic mice to a variety of chemically diverse sweeteners and glucose polymers with three different measures: consumption in 48-h two-bottle preference tests, initial licking responses, and responses of the chorda tympani nerve. The results were generally consistent across the three measures. Allelic variation of the Tas1r3 gene influenced taste responsiveness to nonnutritive sweeteners (saccharin, acesulfame-K, sucralose, SC-45647), sugars (sucrose, maltose, glucose, fructose), sugar alcohols (erythritol, sorbitol), and some amino acids (D-tryptophan, D-phenylalanine, L-proline). Tas1r3 genotype did not affect taste responses to several sweet-tasting amino acids (L-glutamine, L-threonine, L-alanine, glycine), glucose polymers (Polycose, maltooligosaccharide), and nonsweet NaCl, HCl, quinine, monosodium glutamate, and inosine 5'-monophosphate. Thus Tas1r3 polymorphisms affect taste responses to many nutritive and nonnutritive sweeteners (all of which must interact with a taste receptor involving T1R3), but not to all carbohydrates and amino acids. In addition, we found that the genetic architecture of sweet taste responsiveness changes depending on the measure of taste response and the intensity of the sweet taste stimulus. Variation in the T1R3 receptor influenced peripheral taste responsiveness over a wide range of sweetener concentrations, but behavioral responses to higher concentrations of some sweeteners increasingly depended on mechanisms that could override input from the peripheral taste system.


Effect of oleic acid on the viability of different freeze-dried Lactiplantibacillus plantarum strains.
Freeze drying is one of the most convenient ways to preserve microorganisms, but in the freeze-drying process, strains will inevitably suffer varying degrees of damage under different conditions. The deterioration of cell membrane integrity is one of the main forms of damage. The type and ratio of fatty acids in the cell membrane affect its characteristics. Therefore, it is worth investigating whether certain fatty acids can increase freeze-drying resistance. In this study, we found that adding a low concentration of oleic acid to a cryoprotectant could increase survival rate of strains of Lactiplantibacillus plantarum following freeze drying, and the optimal concentration of oleic acid was determined to be 0.001%. When 0.001% oleic acid was added to phosphate-buffered saline, the freeze-drying survival rate of L. plantarum increased by up to 6.63 times. Adding 0.001% oleic acid to sorbitol, the survival rate of L. plantarum increased by as much as 3.65 times. The 0.001% oleic acid-sucrose cryoprotectant resulted in a freeze-drying survival rate of L. plantarum of about 90%, a 2.26-fold improvement compared with sucrose alone. Although the effect of oleic acid depends on the cryoprotectants used and the strain treated, addition of oleic acid showed significant improvement overall. Further experiments showed that adding a low concentration of oleic acid to the cryoprotectants improved the freeze-drying survival rate of L. plantarum by maintaining cell membrane integrity and lactate dehydrogenase activity. Our findings provide a new strategy for safeguarding bacterial viability in commonly used cryoprotectants by the addition of a common food ingredient, which may be extensively applied in the food industry.


Thermal/Near-Infrared Light Dual-Responsive Reversible Two-Way Shape Memory cEVA/2D-MoO2 Composite for Multifunctional Applications.
Light-responsive reversible two-way shape memory polymers (2W-SMPs) are highly promising for many fields due to indirect heating, clean, and remote control. In this work, a composite with both thermal- and near-infrared (NIR) light-induced reversible two-way shape memory effect (2W-SME) is prepared by doping extremely little quantities of 2D non-layered molybdenum dioxide nanosheets (2D-MoO2 ) into semicrystalline poly(ethylene-co-vinyl acetate) (EVA) networks. This is the first report on light-induced reversible two-way shape memory composites employing 2D-MoO2 as photothermal fillers. Upon switching the NIR light on and off, due to the excellent photothermal feature and stability of 2D-MoO2 , the composite exhibits remarkable light-induced reversible 2W-SME. A light-driven actuator for sensing applications is designed based on the composite and the circuit, where the lamp acting as an alarm can raise and fade upon responding to NIR light. A completely flexible, fuel-free self-walking soft robot is designed based on the advantages of the light-responsive reversible 2W-SMPs. Additionally, the composite acting as a light-fueled crane is able to lift and lower a load that is 3846 times its own weight. The results demonstrate that the prepared composite has a promising prospect for applications as actuators, self-walking soft robot and crane.


[Improvement of cognition after trepanation for the chronic subdural hematoma].
The cognitive functions were examined in the pre- and post-operative stages about the 16 cases of the chronic subdural hematoma. The evaluation was performed with MMS (mini-mental state) test. Although the average of the total score was 16.6 in the preoperative stage, it increased to 22.7 in the postoperative stage. Orientation of time and place, recall and the ability of writing a sentence, which were impaired prominently in the preoperative stage, improved significantly in the postoperative stage. The improvement was poorer in the patients with gait instability and in those older than 80 years. Trepanation surgery seems to be efficient for the improvement of cognition.


A Cbx8-containing polycomb complex facilitates the transition to gene activation during ES cell differentiation.
Polycomb proteins play an essential role in maintaining the repression of developmental genes in self-renewing embryonic stem cells. The exact mechanism allowing the derepression of polycomb target genes during cell differentiation remains unclear. Our project aimed to identify Cbx8 binding sites in differentiating mouse embryonic stem cells. Therefore, we used a genome-wide chromatin immunoprecipitation of endogenous Cbx8 coupled to direct massive parallel sequencing (ChIP-Seq). Our analysis identified 171 high confidence peaks. By crossing our data with previously published microarray analysis, we show that several differentiation genes transiently recruit Cbx8 during their early activation. Depletion of Cbx8 partially impairs the transcriptional activation of these genes. Both interaction analysis, as well as chromatin immunoprecipitation experiments support the idea that activating Cbx8 acts in the context of an intact PRC1 complex. Prolonged gene activation results in eviction of PRC1 despite persisting H3K27me3 and H2A ubiquitination. The composition of PRC1 is highly modular and changes when embryonic stem cells commit to differentiation. We further demonstrate that the exchange of Cbx7 for Cbx8 is required for the effective activation of differentiation genes. Taken together, our results establish a function for a Cbx8-containing complex in facilitating the transition from a Polycomb-repressed chromatin state to an active state. As this affects several key regulatory differentiation genes this mechanism is likely to contribute to the robust execution of differentiation programs.


Risk factors for incident asthma and COPD in a cohort of young adults.
The aim of the study was to describe potential shared risk factors for incident asthma and COPD in a population-based, 9-year follow-up study. From a cohort of 1191 individuals, aged 20-44, who participated in baseline survey including spirometry, bronchial challenge, and skin prick test (SPT) 742 subjects (62%) were reexamined at follow-up in 2012-2014. A total of 27 incident cases of asthma and 22 cases of COPD were identified at follow-up corresponding to an incidence rate of 5.8 (95% CI 3.9-8.4) and 3.5 (2.2-5.3) per 1000 person years, respectively. Among the identified COPD cases a total of 12 were Asthma-COPD Overlap Syndrome (ACOS). Atopy defined by positive SPT was a risk factor for asthma in males (OR 7.54; 95% CI 1.24-45.90), whereas risk factors in females were nasal allergy (3.81; 1.20-12.11), FEV1 <100% predicted (3.96; 1.07-14.62) and parental asthma (3.06; 1.00-9.40). Risk factors for COPD in males were bronchial hyperresponsiveness (23.13; 1.41-380.50) and FEV1 <100% predicted (all male cases had FEV1 <100% predicted) and in females current smoking (3.34; 1.16-9.59) and asthma at baseline (5.21; 1.48-18.34). No shared risk factors for incident asthma and COPD were found. Despite low power when stratifying by sex risk factors for incident asthma and COPD emphasize considerable gender differences.


Dendritic cells: a specialized complex system of antigen presenting cells.
The dendritic cell (DC) network is a specialized system for presenting antigen to naive or quiescent T cells, and consequently plays a central role in the induction of T cell and B cell immunity in vivo. Despite considerable achievements in the last ten years, in our understanding of how DC induce and regulate immune responses, much remains to be learned about this complex system of cells. The history and current status of DC termed "directors of the immune system orchestra" is reviewed. The present understanding of DC cell biology, function and use, taking into account their complexity is discussed.


Increased N,N-Dimethyl Leucine Isobaric Tag Multiplexing by a Combined Precursor Isotopic Labeling and Isobaric Tagging Approach.
Multiplex isobaric tags have become valuable tools for high-throughput quantitative analysis of complex biological samples in discovery-based proteomics studies. Hybrid labeling strategies that pair stable isotope mass difference labeling with multiplex isobaric tag-based quantification further facilitate these studies by greatly increasing multiplexing capability. In this work, we present a cost-effective chemical labeling approach that couples duplex stable isotope dimethyl labeling with our custom 12-plex N,N-dimethyl leucine (DiLeu) isobaric tags in a combined precursor isotopic labeling and isobaric tagging (cPILOT) strategy that is compatible with a wide variety of biological samples and permits 24-plex quantification in a single LC-MS/MS experiment. We demonstrate the utility of the DiLeu cPILOT approach by labeling yeast digests and performing proof-of-principle quantification experiments on the Orbitrap Fusion Lumos.


Chorea and Orofaciolingual Dystonia in a 40 Year Old Male.
Neuroacanthocytosis is a heterogeneous group of disorders which result in progressive neurodegeneration, predominantly of the basal ganglia, and erythrocyte acanthocytosis. We report a case of neuroacanthocytosis with typical phenotype of choreoacanthocytosis. A 40 year male presented with features of chorea with orofaciolingual dystonia producing eating and speech difficulties. There were features of self mutilation in form of lip and tongue biting. Peripheral blood smear examination revealed acanthocytes in our patient. Neuroimaging showed bilateral caudate atrophy and nerve conduction study showed motor axonal neuropathy. This case report describes the typical features and investigations to diagnose this rare disorder which is usually underdiagnosed.


Effect of formulation on the stability and aerosol performance of a nebulized antibody.
Most monoclonal antibodies (mAbs) are administered to patients intravenously to ensure high bioavailability as rapidly as possible. The airways, however, are an attractive delivery route for mAbs for the treatment of lung diseases, making it possible to increase their concentration in the target organ while limiting their systemic passage. Several challenges must be overcome for translation into clinical practice. For example, the drug and device must be paired for the efficient and reliable deposition of a pharmacologically active and safe mAb in the lung region of interest. Mesh nebulizers appear to be the most effective aerosol-producing devices for delivering large amounts of biopharmaceutical while limiting protein instability during nebulization. We used metrological and analytic methods to analyze the effect of both antibody concentration and surfactant addition on aerosol performance and antibody integrity. These two factors had a limited effect on aerosol performance, but affected antibody aggregation. The addition of surfactants to antibody formulations at concentrations appropriate for lung administration markedly reduced the formation of medium or large aggregates, as shown by dynamic light scattering and fluorescence microscopy. Aggregation was also dependent on the type of mesh nebulizer, highlighting the need to optimize drug and device together.


Kinetics of cell proliferation and polyamine synthesis during Ehrlich ascites tumor growth.
The kinetics of cell proliferation and polyamine synthesis during Ehrlich ascites tumor growth were studied. The steady deceleration of the specific growth rate with increasing tumor mass that was observed was attributable to a prolongation of the cell cycle, particularly of the S and G2 phases. The cell cycle time (Tc) was 43.3 hr (TG1 equals 10.8, TS equals 26.8, and TG2 equals 5.7 hr) on the seventh day of growth and 76.0 hr (TG1 equals 14.0, TS equals 52.0, and TG2 equals 10.0 hr) on the tenth day of growth. The growth fraction showed a decrease from 0.77 to 0.60 during the 7- to 10-day tumor growth interval. The cell death rate remained low and essentially unchanged during this period. A high correlation was found between polyamine synthesis (ornithine decarboxylase activity) and the specific growth rate; the correlation coefficient was 0.985. There was also a high positive correlation between the cellular polyamine (spermidine and spermine) and nucleic acid content (spermidine: DNA equals 0.916, spermine: DNA equals 0.947, spermidine:RNA equals 0.907, and spermine: RNA equals 0.881). These observations suggest that there may be a functional coupling between polyamines and nucleic acids, and they support the hypothesis that polyamines play an important role in DNA replication and cell division.


Rural Maternal Health Care Outcomes, Drivers, and Patient Perspectives.
Rural communities are a vital segment of the US population; however, these communities are shrinking, and their population is aging. Rural women experience health disparities including increased risk of maternal morbidity and mortality. In this article, we will explore these trends and their determinants both within and external to the health care system. Health care providers, public health professionals, and policymakers should be aware of these social and structural factors that influence health outcomes and take action to reduce generational cycles of health disparity. Opportunities to improve the health and pregnancy outcomes for rural women and rural populations are highlighted.


Direct measurement of bacterial penetration through surgical gowns: a new method.
There is a paucity of data on the penetration of bacteria through surgical gowns during operations. A simple new method was developed, using Petri dishes filled with horse blood agar that were attached to the outside of the gown material. This was used to assess bacterial penetration through disposable spun-bonded polyester gowns and re-usable woven polyester gowns during normal use. There was a significant difference between the two gown types when tested in the axilla (P = 0.02), the groin (P = 0.02) and the peri-anal region (P < 0.01), with the disposable gowns performing to a higher standard. Re-usable gowns demonstrated variation in penetrability, and for this reason, may be unsuitable for use in orthopaedic implant surgery.


A lineage-specific gene encoding a major matrix protein of the sea urchin embryo spicule. II. Structure of the gene and derived sequence of the protein.
A lambda gt11 cDNA clone isolated by use of a polyclonal antispicule matrix protein antiserum is shown in the accompanying paper [S. C. Benson, H. M. Sucov, L. Stephens, E. H. Davidson, and F. Wilt (1987) Dev. Biol. 120, 499-506] to encode a prominent 50-kDa spicule matrix protein (SM50). This clone was used to select homologous genomic recombinants, and the structure of the gene was determined. The SM50 gene occurs once per haploid genome. It contains a single intron located within the 35th codon. A unique transcription initiation site 110 nucleotide pairs prior to the translation start signal was mapped by primer extension. The mRNA is 1895 nucleotides in length, excluding the 3' poly(A) sequence, and contains a single open reading frame 450 codons in length. Though rare in whole embryo RNA the prevalence of the SM50 mRNA is calculated to be about 1% of the total mRNA in skeletogenic mesenchyme cells. The derived peptide sequence indicates a typical N-terminal signal peptide, and an N-linked glycosylation site near the C terminus. About 45% of the length of the protein is included in a domain composed of consecutive approximate repetitions of a 13-amino-acid element, the consensus sequence of which is Trp-Val-Gly-Asp-Asn-Gln-Ala-LeuTrp-Val-IleAsp-Asn-GlnPro+ ++-ValGlu. The protein also contains an internal domain unusually rich in proline residues and a very basic C-terminal region.


Unique Challenges to Diagnosing Human Herpesvirus-6 (HHV-6) Encephalitis Following Post-Hematopoietic Stem Cell Transplant: A Case and Brief Review.
A patient with an ultimate diagnosis of human herpesvirus-6 (HHV-6) encephalitis developed central nervous system (CNS) symptoms 13 days after undergoing myeloablative haploidentical allogeneic hematopoietic stem cell transplant (HSCT). Due to the patient's body habitus, magnetic resonance (MR) imaging was not obtained until the onset of retrograde amnesia on day +24. MR imaging and other clinical findings eliminated all skepticism of HHV-6 encephalitis and HHV-6 antivirals were initiated on day +28, leading to gradual recovery. This case demonstrates some of the factors that may complicate the diagnosis of post-alloHSCT HHV-6 encephalitis. Because HHV-6 encephalitis and viremia can occur without warning, a single negative study should not exclude future development, especially if CNS symptoms are present. Acute graft-versus-host disease and cord blood transplantation are both significant risk factors for HHV-6 encephalitis. Human leukocyte antigen (HLA) mismatch, engraftment complications, or certain HLA alleles have also been associated with HHV-6 encephalitis. Chromosomally integrated HHV-6 must also be ruled out to prevent inappropriate and potentially harmful administration of antivirals. Due to the severe short- and long-term sequelae of HHV-6 encephalitis, appropriate treatment should be administered as soon as possible.


First report of the post-fire morel Morchella exuberans in eastern North America.
Reports of true morels (Morchella) fruiting on conifer burn sites are common in western North America where five different fire-adapted species of black morels (Elata Clade) have been documented based on multilocus phylogenetic analyses. Fruiting of post-fire morels in eastern North America, by comparison, are rare and limited to a report from Minnesota in 1977 and eastern Ontario in 1991. Here, nuc rDNA internal transcribed spacer (ITS1-5.8S-ITS2 = ITS) sequences were used to identify the post-fire morel that fruited in great abundance the year following the 2012 Duck Lake Fire in the Upper Peninsula of Michigan and after the 2016 large-scale fire in the Great Smoky Mountains National Park in Tennessee as M. exuberans. A preliminary phylogenetic analysis suggests that the collections from eastern North America may be more closely related to those from Europe than from western North America, Europe, and China.


The Bacillus subtilis ureABC operon.
The Bacillus subtilis ureABC operon encodes homologs of the three subunits of urease enzymes of the family Enterobacteriaceae. Disruption of ureC prevented utilization of urea as a nitrogen source and resulted in a partial growth defect in minimal medium containing limiting amounts of arginine or allantoin as the sole nitrogen source.


[Epidemiology of imported malaria and entomological study of breeding sites of potential risk areas in the province of Khemisset (Morocco)].
No case of autochthonous malaria has been detected in Morocco since 2004. This achievement is due to a national strategy to combat the disease by appropriate and well-organized disease detection and treatment, as well as control of the mosquito vector of the disease, the female Anopheles mosquito. Nonetheless, imported malaria cases have been increasing (75 in 2007), due to the rise in international travel and migration from countries where the disease is endemic. This work is divided into two parts: the first part is a retrospective study of the cases of imported malaria identified by optical microscopy in the Laboratory of Medical Entomology of the Khemisset Provincial Delegation of Health from 2000 to 2010. The second part is an entomological study conducted in 2010 of the Culicidae insect family, especially the Anopheles genus. The results show that of 176,457 requests for parasite testing, 14 were positive. All positive samples came from men older than 23 years. The cases identified are imported from two African countries: the Democratic Republic of Congo (86%) and Ivory Coast (14%). The years of high incidence were 2003 with four cases and 2000, 2004 and 2006 with two cases. The species found are Plasmodium falciparum in 13 cases (93%) and Plasmodium ovale in one case (7%). The vector of autochthonous malaria, which was eliminated in 2004, is Anopheles (Anopheles) labranchiae Fallerouni 1926, and it was the dominant species found in our entomological study (424 larvae). The other species were found in breeding sites in potential at-risk locations in the study area.


A small-molecule IRF3 agonist functions as an influenza vaccine adjuvant by modulating the antiviral immune response.
Vaccine adjuvants are essential to drive a protective immune response in cases where vaccine antigens are weakly immunogenic, where vaccine antigen is limited, or where an increase in potency is needed for a specific population, such as the elderly. To discover novel vaccine adjuvants, we used a high-throughput screen (HTS) designed to identify small-molecule agonists of the RIG-I-like receptor (RLR) pathway leading to interferon regulatory factor 3 (IRF3) activation. RLRs are a group of cytosolic pattern-recognition receptors that are essential for the recognition of viral nucleic acids during infection. Upon binding of viral nucleic acid ligands, the RLRs become activated and signal to transcription factors, including IRF3, to initiate an innate immune transcriptional program to control virus infection. Among our HTS hits were a series of benzothiazole compounds from which we designed the lead analog, KIN1148. KIN1148 induced dose-dependent IRF3 nuclear translocation and specific activation of IRF3-responsive promoters. Prime-boost immunization of mice with a suboptimal dose of a monovalent pandemic influenza split virus H1N1 A/California/07/2009 vaccine plus KIN1148 protected against a lethal challenge with mouse-adapted influenza virus (A/California/04/2009) and induced an influenza virus-specific IL-10 and Th2 response by T cells derived from lung and lung-draining lymph nodes. Prime-boost immunization with vaccine plus KIN1148, but not prime immunization alone, induced antibodies capable of inhibiting influenza virus hemagglutinin and neutralizing viral infectivity. Nevertheless, a single immunization with vaccine plus KIN1148 provided increased protection over vaccine alone and reduced viral load in the lungs after challenge. These findings suggest that protection was at least partially mediated by a cellular immune component and that the induction of Th2 and immunoregulatory cytokines by a KIN1148-adjuvanted vaccine may be particularly beneficial for ameliorating the immunopathogenesis that is associated with influenza viruses.


Chronic mountain sickness: recent studies of the relationship between hemoglobin concentration and oxygen transport.
Although an increase in hemoglobin concentration [Hb] in high altitude residents assists oxygen transport, excessive polycythemia ([Hb] > or = 21 g/100 mL) may cause the syndrome of chronic mountain sickness (CMS). A recent theoretical analysis has suggested that increasing [Hb] above 18 g/100 mL provides no further benefit in oxygen transport at rest. To test this hypothesis, we examined oxygen transport at rest for given arterial oxygen saturations (Sa(O2), in classes at intervals of 5%) as reported in 206 residents of various altitudes. For Sa(O2) of 97% versus 87%, [Hb] and a-v oxygen content difference increased (respectively, 14.5 to 17.5 g/100 mL and 4.11 to 5.03 volume %). As Sa(O2) fell further to 66%, a-v progressively decreased to 3.77 volume %, despite an increase in [Hb] to 24.2 g/100 mL. Over the Sa(O2) range of 97% to 66%, the a-v difference changed little (-8%) compared to other subjects made acutely hypoxic (-33%), for Sa(O2) change from 97% to 75%. The results suggest that increasing [Hb] allows greater oxygen extraction (a cardiac output sparing effect), which is maximal at Sa(O2) of 87% and a [Hb] of 17.5 g/100 mL. For more severe hypoxemia, even to Sa(O2) of 66%, both increasing [Hb] and increasing output are utilized for oxygen transport.


General practitioners' attitudes and ethical decisions in end-of-life care after a year of interactive Internet-based training.
Personal characteristics affect physicians' end-of-life decisions. Education in palliative care (PC) faces a challenge in influencing doctors' attitudes and decision making in PC. A one-year Internet-mediated education project was arranged for general practitioners in Finland to provide postgraduate education in PC. A questionnaire was sent before and after the project to the education group (EG, n = 79) and a control group (CG, n = 100). Treatment decisions for presented scenarios and attitudes toward PC were elicited. EG doctors had previously participated in postgraduate training in PC more often than had CG doctors (93% vs 29%). For a young terminal cancer patient, EG doctors chose CPR less often than did CG doctors (7% vs 33%, p = 0.004); 30% of EG doctors would have continued antibiotic treatment in the case of a terminal cancer patient, vs 55% of CG doctors (p = 0.023). EG doctors more often agreed with the statement "Terminal care is satisfying" (p < 0.001). Treatment decisions or attitudes did not change significantly in either group during the year, the EG doctors were more satisfied with their work after the project. There are substantial differences in decision making between those who have experience and special interest in the subject and those who do not. To achieve the goals of education in PC, the most difficult target group, the "silent majority" of doctors, should be reached.


A novel nickel avidin-biotin-peroxidase method for histochemical visualization of neurofibrillary tangles, senile plaques, and neuropil threads.
A reliable new method was developed for detecting neurofibrillary tangles, senile plaques, and neuropil threads in human neural tissue. Excellent morphological definition of the pathological structures was achieved with this procedure without staining normal neuronal and glial elements. The technique was applied to cortical tissue from eight patients with Alzheimer's disease and three controls. Histological sections from these cases were incubated in an avidin-biotin-peroxidase complex solution containing 0.5% nickel ammonium sulfate, followed by visualization in 3,3'-diaminobenzidine and H2O2. Although the avidin-biotin system is routinely used in immunohistochemistry, no antibodies were employed in the present procedure. This technique has advantages over silver impregnation methods because it requires very little monitoring of critical steps and yields superior results. The method is suitable for processing large numbers of tissue sections per staining run, and the results are highly reproducible. These features are advantageous in research studies comparing the distribution of lesions in large numbers of cases. The precise mechanism of staining has not been determined; however, conditions such as nickel concentrations, pH, and avidin-biotin-peroxidase complex concentrations were varied to examine critical steps in the process.


Laser-capture microdissection: applications in routine molecular dermatopathology.
Advances in molecular pathology with the introduction of the Southern blot technique and the polymerase chain reaction (PCR) have emerged as important tools, which are frequently used in routine dermato-histopathology. Applications for PCR-based diagnostics are particularly helpful for the determination of clonality in cutaneous lymphocytic infiltrates and for detection of infectious agents, such as herpes simplex virus (HSV), varicella zoster virus (VZV), Borrelia burgdorferi, Mycobacteria, Leishmania, and Treponema pallidum. As biopsies are always composed of different cells, the cells of interest are often only a minor population. As a consequence, their specific DNA is diluted by the majority of contaminating cells. Another problem is the time- and labor-intensive DNA extraction, because usually only formalin-fixed, paraffin-embedded tissue is available, which makes molecular diagnostics a time and labor consuming, and consequently a cost-intensive procedure. To overcome these shortcomings and to eventually shorten the time to generate a result, we introduce a laser-capture microdissection (LCM)-based method for the detection of infectious agents and clonality. Only the cells of interest for the particular indication are microdissected (e.g. epidermal cells for HSV and VZV and lymphocytes for clonality analysis) and subjected to PCR amplification. Due to an accelerated DNA-extraction procedure which generates DNA in 5 h (compared to 3-4 days using conventional DNA extraction), we are able to generate a result within one working day.


Growth and genetic stability of the ts-1 mutant of respiratory syncytial virus at restrictive temperatures.
An in vitro study was performed to define in greater detail those factors which favored the growth of the ts-1 mutant of respiratory syncytial virus under restrictive conditions and the emergence of genetically altered virus with decreased temperature sensitivity. Replication of ts-1 occurred at each of the restrictive temperatures of 37, 38, and 39 C, even through plaque formation was not observed. The level of virus growth under restrictive conditions was inversely related to the incubation temperature and directly related to the multiplicity of infection. These relationships appeared to reflect the effect of restrictive temperature in reducing the quantity of virus produced and released from an infected cell. Under restrictive conditions the production of genetically altered virus which exhibited reduced temperature sensitivity was directly related to the multiplicity of infection and inversely related to temperature. Production of genetically altered virus was not observed under permissive conditions.


Role of matrix in an early postentry step in the human immunodeficiency virus type 1 life cycle.
The matrix protein of human immunodeficiency virus type 1 (HIV-1) has been reported to play a crucial role in the targeting of the Gag polyprotein precursor to the plasma membrane and in the incorporation of viral envelope glycoproteins into budding virions. In this report, we present evidence that mutation of a highly conserved Leu at matrix amino acid 20 blocks or markedly delays virus replication in a range of cell types, including T-cell lines, primary human peripheral blood mononuclear cells, and monocyte-derived macrophages. These mutations do not impair virus assembly and release, RNA encapsidation, or envelope glycoprotein incorporation into virions but rather cause significant defects in an early step in the virus life cycle, as measured by single-cycle infectivity assays and the analysis of viral DNA synthesis early postinfection. This infectivity defect is independent of the type of envelope glycoprotein carried on mutant virions; similar results are obtained in pseudotyping experiments using wild-type or truncated HIV-1 envelope glycoproteins, the amphotropic murine leukemia virus envelope, or the vesicular stomatitis G protein. Intriguingly, matrix residue 20 mutations also increase the apparent binding of Gag to membrane, accelerate the kinetics of Gag processing, and induce defects in endogenous reverse transcriptase activity without affecting virion density or morphology. These results help elucidate the function of matrix in HIV-1 replication.


Finding the flaw.
midwives, practice nurses, health visitors and school nurses can provide health education which is essential in preventing incontinence. Minority ethnic groups must not be forgotten--leaflets are available in a variety of languages. only by identifying the type and cause of incontinence can appropriate treatment and management be decided. Practice nurses and health visitors can be involved in assessment. if more than occasional light incontinence is identified or the practice nurse or health visitor does not feel competent to deal with the problem, women must be encouraged to seek further help and should be referred to their GP or local continence advisory service, which usually takes self-referrals.


Maternal depression and early positive parenting predict future conduct problems in young children with attention-deficit/hyperactivity disorder.
Children with attention-deficit/hyperactivity disorder (ADHD) are at risk for adverse outcomes such as substance abuse and criminality, particularly if they develop conduct problems. Little is known about early predictors of the developmental course of conduct problems among children with ADHD, however. Parental psychopathology and parenting were assessed in 108 children who first met Diagnostic and Statistical Manual of Mental Disorders (4th ed.) criteria for ADHD at 4-7 years old. When demographic variables and baseline ADHD and conduct problems were controlled, maternal depression predicted conduct problems 2-8 years following the initial assessment, whereas positive parenting during the structured parent- child interaction task predicted fewer future conduct problems. These findings suggest that maternal depression is a risk factor, whereas early positive parenting is a protective factor, for the developmental course of conduct problems among children with ADHD.


Genome-wide study of gene variants associated with differential cardiovascular event reduction by pravastatin therapy.
Statin therapy reduces the risk of coronary heart disease (CHD), however, the person-to-person variability in response to statin therapy is not well understood. We have investigated the effect of genetic variation on the reduction of CHD events by pravastatin. First, we conducted a genome-wide association study of 682 CHD cases from the Cholesterol and Recurrent Events (CARE) trial and 383 CHD cases from the West of Scotland Coronary Prevention Study (WOSCOPS), two randomized, placebo-controlled studies of pravastatin. In a combined case-only analysis, 79 single nucleotide polymorphisms (SNPs) were associated with differential CHD event reduction by pravastatin according to genotype (P<0.0001), and these SNPs were analyzed in a second stage that included cases as well as non-cases from CARE and WOSCOPS and patients from the PROspective Study of Pravastatin in the Elderly at Risk/PHArmacogenomic study of Statins in the Elderly at risk for cardiovascular disease (PROSPER/PHASE), a randomized placebo controlled study of pravastatin in the elderly. We found that one of these SNPs (rs13279522) was associated with differential CHD event reduction by pravastatin therapy in all 3 studies: P = 0.002 in CARE, P = 0.01 in WOSCOPS, P = 0.002 in PROSPER/PHASE. In a combined analysis of CARE, WOSCOPS, and PROSPER/PHASE, the hazard ratio for CHD when comparing pravastatin with placebo decreased by a factor of 0.63 (95% CI: 0.52 to 0.75) for each extra copy of the minor allele (P = 4.8 × 10(-7)). This SNP is located in DnaJ homolog subfamily C member 5B (DNAJC5B) and merits investigation in additional randomized studies of pravastatin and other statins.


How pharmacological receptor theory can guide new drug discovery CCR5 HIV inhibitors in AIDS as a case study.
The simple concept of quantifying drug activity through measurement of affinity and efficacy is challenged by the discovery, through new assay technologies, that ligands may have different 'efficacies' for different receptor-driven cellular processes. Thus, a ligand may directly produce the complete behavior pattern of the endogenous natural agonist or only portions of this pattern; the challenge is to quantify these differences in ligands and identify the therapeutically useful phenotypes. Another effect, related to the allosteric nature of the 7-transmembrane domain superfamily of receptors, is the ability of molecules to co-bind to receptors (like antagonists) and selectively modify the effects of endogenous agonists, in essence to filter effects and produce a different pattern of signaling. One realm where this may be useful is the blockade of pathologically related effects of receptors that otherwise are needed for normal physiology. These concepts are illustrated in the context of new allosteric CCR5 HIV-1 entry inhibitors. The allosteric properties of probe dependence and texture in antagonism are related to potential advantages in sparing of normal physiological function and reduction in susceptibility to viral resistance.


Five classic articles in public health.
In this brief review, Dr. Jonathan Borak comments on five seminal papers that helped shape the fields of epidemiology and public health. These papers include Hill's criteria for inferring causality; the first proof of the multistage theory of cancer; the first evidence that subclinical lead exposures can cause neurobehavioral impairment in children; a simple yet robust study that had a major influence on setting current air pollution policies; and a landmark review of the general public's perception of risk in relation to actual public health hazard.


High-temperature cultivation of recombinant Pichia pastoris increases endoplasmic reticulum stress and decreases production of human interleukin-10.
The yeast Pichia pastoris (P. pastoris) has become a popular 'cell factory' for producing heterologous proteins, but production widely varies among proteins. Cultivation temperature is frequently reported to significantly affect protein production; however, the underlying mechanisms of this effect remain unclear. A P. pastoris strain expressing recombinant human interleukin-10 (rhIL-10) under the control of the AOX1 promoter was used as the model in this study. This system shows high-yield rhIL-10 production with prolonged methanol-induction times when cultured at 20°C but low-yield rhIL-10 production and higher cell death rates when cultured at 30°C. Further investigation showed that G3-pro-rhIL10, an immature form of rhIL-10 that contains the glycosylation-modified signal peptide, remained in the ER for a prolonged period at 30°C. The retention resulted in higher ER stress levels that were accompanied by increased ROS production, Ca(2+) leakage, ER-containing autophagosomes, shortened cortical ER length and compromised induction of the unfolded protein response (UPR). In contrast, G3-pro-rhIL10 was quickly processed and eliminated from the ER at 20°C, resulting in a lower level of ER stress and improved rhIL-10 production. High-temperature cultivation of an rhIL-10 expression strain leads to prolonged retention of immature G3-pro-rhIL10 in ER, causing higher ER stress levels and thus greater yeast cell death rates and lower production of rhIL-10.


Dual dean entrainment with volume ratio modulation for efficient droplet co-encapsulation: extreme single-cell indexing.
The future of single cell diversity screens involves ever-larger sample sizes, dictating the need for higher throughput methods with low analytical noise to accurately describe the nature of the cellular system. Current approaches are limited by the Poisson statistic, requiring dilute cell suspensions and associated losses in throughput. In this contribution, we apply Dean entrainment to both cell and bead inputs, defining different volume packets to effect efficient co-encapsulation. Volume ratio scaling was explored to identify optimal conditions. This enabled the co-encapsulation of single cells with reporter beads at rates of ∼1 million cells per hour, while increasing assay signal-to-noise with cell multiplet rates of ∼2.5% and capturing ∼70% of cells. The method, called Pirouette coupling, extends our capacity to investigate biological systems.


Polymorphisms in the yeast galactose sensor underlie a natural continuum of nutrient-decision phenotypes.
In nature, microbes often need to "decide" which of several available nutrients to utilize, a choice that depends on a cell's inherent preference and external nutrient levels. While natural environments can have mixtures of different nutrients, phenotypic variation in microbes' decisions of which nutrient to utilize is poorly studied. Here, we quantified differences in the concentration of glucose and galactose required to induce galactose-responsive (GAL) genes across 36 wild S. cerevisiae strains. Using bulk segregant analysis, we found that a locus containing the galactose sensor GAL3 was associated with differences in GAL signaling in eight different crosses. Using allele replacements, we confirmed that GAL3 is the major driver of GAL induction variation, and that GAL3 allelic variation alone can explain as much as 90% of the variation in GAL induction in a cross. The GAL3 variants we found modulate the diauxic lag, a selectable trait. These results suggest that ecological constraints on the galactose pathway may have led to variation in a single protein, allowing cells to quantitatively tune their response to nutrient changes in the environment.


[Homograft in the therapy of cardiovascular diseases].
Despite considerable progress in prosthetic valve technology, there is actually no ideal artificial heart valve that may be employed in all circumstances, when replacement of a diseased aortic valve is necessary. The choice for optimal valve substitute includes mechanical prosthesis, bioprosthetic xenografts, homografts and pulmonary autografts. More recently an aortic valve sparing operation has been proposed in younger patients presenting with anulo-aortic ectasia. The use of homograft heart valves has now been widely accepted in the treatment of congenital heart defects, as well as for an increasing number of valvular pathologies, especially in infective endocarditis. Heart valve preservation by cryopreservation techniques helps to store the small amount of donor material for special indications without loss of quality, thus permitting elective surgery. In vascular surgery, some concern persists regarding the use of prosthetic material for the treatment of mycotic aneurysms and graft infection. In situ repair with a new vascular prosthesis and resection followed by extra-anatomic reconstruction carry a high peri-operative mortality and substantial morbidity. Cryopreserved vascular homografts represent a valuable alternative in these challenging situations.


Comparative studies of genes encoding thermostable L-2-halo acid dehalogenase from Pseudomonas sp. strain YL, other dehalogenases, and two related hypothetical proteins from Escherichia coli.
We have determined the nucleotide sequence of the gene encoding thermostable L-2-halo acid dehalogenase (L-DEX) from the 2-chloroacrylate-utilizable bacterium Pseudomonas sp. strain YL. The open reading frame consists of 696 nucleotides corresponding to 232 amino acid residues. The protein molecular weight was estimated to be 26,179, which was in good agreement with the subunit molecular weight of the enzyme. The gene was efficiently expressed in the recombinant Escherichia coli cells: the amount of L-DEX corresponds to about 49% of the total soluble proteins. The predicted amino acid sequence showed a high level of similarity to those of L-DEXs from other bacterial strains and haloacetate dehalogenase H-2 from Moraxella sp. strain B (38 to 57% identity) but a very low level of similarity to those of haloacetate dehalogenase H-1 from Moraxella sp. strain B (10%) and haloalkane dehalogenase from Xanthobacter autotrophicus GJ10 (12%). By searching the protein amino acid sequence database, we found two E. coli hypothetical proteins similar to the Pseudomonas sp. strain YL L-DEX (21 to 22%).


High assimilation of the sacrum in a sample of American skeletons: prevalence, pelvic size, and obstetrical and evolutionary implications.
High assimilation sacrum is fusion of the caudal-most lumbar vertebra to the first sacral vertebra. Previous studies have shown that high assimilation is associated with clinical problems, including obstetrical difficulty. This study used adult American males (n = 1,048) and females (n = 1,038) of the Hamann-Todd and Terry skeletal collections to determine the prevalence of high assimilation and its effect on pelvic size, and to consider the obstetrical and evolutionary implications of high assimilation. The prevalence of high assimilation in this sample is 6.3%, with males and females not differing significantly from one another in their prevalence. This prevalence is near the median for that reported in 41 other samples. In both males and females, individuals with high assimilation have significantly longer anteroposterior and posterior sagittal diameters of the inlet, and shorter sacrum compared to those with a nonassimilated sacrum. Females with high assimilation have a significantly narrower sacral angulation (i.e., reduced inclination of ventral axis of sacrum), and shorter posterior sagittal diameter of the outlet compared to those with a nonassimilated sacrum. A short posterior sagittal diameter of the outlet is associated with childbirth difficulty. As high assimilation is partial homeotic transformation of a lumbar vertebra, this study supports previous research that homeotic transformation of vertebrae is selectively disadvantageous.


Exploring spin symmetry-breaking effects for static field ionization of atoms: Is there an analog to the Coulson-Fischer point in bond dissociation?
Löwdin's symmetry dilemma is an ubiquitous issue in approximate quantum chemistry. In the context of Hartree-Fock (HF) theory, the use of Slater determinants with some imposed constraints to preserve symmetries of the exact problem may lead to physically unreasonable potential energy surfaces. On the other hand, lifting these constraints leads to the so-called broken symmetry solutions that usually provide better energetics, at the cost of losing information about good quantum numbers that describe the state of the system. This behavior has previously been extensively studied in the context of bond dissociation. This paper studies the behavior of different classes of HF spin polarized solutions (restricted, unrestricted, and generalized) in the context of ionization by strong static electric fields. We find that, for simple two electron systems, unrestricted Hartree-Fock (UHF) is able to provide a qualitatively good description of states involved during the ionization process (neutral, singly ionized, and doubly ionized states), whereas RHF fails to describe the singly ionized state. For more complex systems, even though UHF is able to capture some of the expected characteristics of the ionized states, it is constrained to a single Ms (diabatic) manifold in the energy surface as a function of field intensity. In this case, a better qualitative picture can be painted by using generalized Hartree-Fock as it is able to explore different spin manifolds and follow the lowest solution due to lack of collinearity constraints on the spin quantization axis.


First cases of animal diseases published in 2005.
In 2003, Veterinary Quarterly started with the publication of a series of articles reviewing reports on "first cases" of animal diseases. Till now six articles have been published. They were arranged in order of animal species. Each article concerned only one species. Starting at this seventh publication, each article discusses "first cases" on more than one animal species. In this article 15 cases published in 2005 are discussed: one in birds, two in cats, three in cattle, four in dogs, two in goats, one in hares, and two in horses. Of each case a short introduction, the bibliographical data of the publication, the abstract of the author(s) (except one case) and some additional information (mainly) derived from the article are given.


Health Profiles of Allied Health Students Enrolled in a Faith-Based University.
The purpose of this study was to explore the health profiles of students at a faith-based university. Data were collected using a modified version of the Health Lifestyle Enhancement Profile (HELP) and HELP-Screener. Participants were 323 undergraduate and graduate allied health students, of whom 64% were female and 36% male, with ages ranging from 19 to 51 years. Associations between variables were explored and a two-step cluster analysis was applied to the data. Results showed that 34.1% of students perceived their health as excellent and 57.2% as good. It was also detected that high ratings of health were positively correlated with high ratings of happiness. Further, students who reported frequent exercise and a healthy diet scored more favorably across all measures of health in comparison to those who reported less frequent exercise and poorer dietary habits. Descriptive analysis revealed that 46.6% of students identified having at least one health problem, with back and neck pain being the most prevalent. Less than half of the study sample reported monitoring their health regularly, avoiding unhealthy foods, avoiding sedentary activities, and engaging in community activities. Considerations for future university-based interventions to support students' health at faith-based institutions are indicated.


The Wnt/β-catenin signaling pathway inhibits osteoporosis by regulating the expression of TERT: an in vivo and in vitro study.
Our study was performed to investigate whether the Wingless and int-1 (Wnt) signaling pathway promotes osteogenic differentiation and inhibits apoptosis in bone marrow mesenchymal stem cells (BMSCs) by regulating telomerase reverse transcriptase (TERT) expression. An in vivo model of osteoporosis (OP) in C57BL/6J mice by bilateral ovariectomy (OVX) and an in vitro model of H2O2-induced BMSCs were established separately. Western blotting was used to detect the expression of the pathway-related proteins TERT, β-catenin, and phosphorylated-glycogen synthase kinase-3beta (p-GSK3β)/GSK3β, the osteogenic-related markers osteopontin (OPN), bone morphogenetic protein 2 (BMP2), and runt-related transcription factor 2 (Runx2), and the apoptosis-related indicators B-cell lymphoma-2 (Bcl-2) and BAX. Osteoblastic phenotypes were also evaluated by alkaline phosphatase (ALP) staining and serum ALP activity assays. Osteogenic differentiation phenotypes in mice were verified by H&E staining, micro-CT, and parameter analysis of the femur. Western blotting results showed that the expression of the pathway-related proteins TERT, β-catenin, p-GSK3β/GSK3β was reduced in OVX mice and H2O2-induced BMSCs, accompanied by downregulated protein expression of osteogenic-related markers and antiapoptotic indicators and upregulated protein expression of apoptotic proteins compared to those in the control group. Mechanistic studies showed that the activation of Wnt signaling pathway in BMSCs promoted β-catenin translocation to the nucleus, as verified by immunofluorescence and facilitated colocalization between β-catenin and TERT, as verified by double-labeling immunofluorescence, thereby promoting osteogenic differentiation and reducing apoptosis. In summary, our experiments confirmed that the GSK3β/β-catenin/TERT pathway could regulate the osteogenic differentiation and apoptosis of BMSCs and that TERT might be a promising target for the future treatment of osteoporosis.


Surgery Start Time Does Not Impact Outcome in Elective Cardiac Surgery.
Determine if surgery start time impacts patient outcomes in elective cardiac surgery. This was a retrospective study. This study was based at a single academic institution. Patients undergoing elective cardiac surgery over a 3-year period were included. There were no interventions. The authors performed a retrospective study of patients undergoing elective cardiac surgery over a 3-year period. They divided their patient groups into those who had an anesthesia start time between 6:00 a.m. and 4:00 p.m. and those who had an anesthesia start time between 4:01 p.m. and 5:59 a.m. In the original sample and propensity-score-matched groups, the authors examined the effects of start time on morbidity, mortality, and several metrics of hospital length of stay. The start time of elective cardiac surgery did not have a statistically significant effect upon mortality, individual or composite morbidity, or hospital length of stay in either the original sample or the propensity-score-matched sample. The authors' results suggested that elective cardiac surgery may be performed late at night without adverse effects, although institutional support for this effort (such as 24-hour intensivist coverage to facilitate fast-track extubation) may have been integral to their findings.


[Clinical value of droplet digital PCR in rapid diagnosis of invasive fungal infection in neonates].
To evaluate the clinical value of droplet digital PCR (ddPCR) in rapid and accurate diagnosis of invasive fungal infection (IFI) in neonates. The highly conserved sequence of fungi 18S RNA was selected as the target sequence, and primers were designed to establish a ddPCR fungal detection system. Blood samples were collected from 83 neonates with high-risk factors for IFI and/or related clinical symptoms in the neonatal intensive care unit (NICU) of a hospital in Shenzhen, China. Blood culture and ddPCR were used for fungal detection. The ddPCR fungal detection system had a specificity of 100% and a sensitivity of 3.2 copies/μL, and had a good reproducibility. Among the 22 blood samples from neonates with a confirmed or clinical diagnosis of IFI, 19 were detected positive by ddPCR. Among the 61 blood samples from neonates who were suspected of IFI or had no IFI, 2 were detected positive by ddPCR. The ddPCR technique can be used for the detection of neonatal IFI and is a promising tool for the screening and even diagnosis of neonatal IFI.


[Drainage for the control of complications after extraction of impacted mandibular third molar: a systematic review].
To assess the efficacy and safety of drainage for the control of the complications following impacted mandibular third molar extraction. To retrieve randomized controlled trials assessing the efficacy and safety of drainage for the control of the complications following impacted mandibular third molar extraction, bibliographic databases including MEDLINE, Cochrane Controlled Trials Register, EMBASE, OPEN SIGLE and China Biology Medicine Database (CBM) were searched on August 23th 2011. References of the included studies and Chinese dental journals were hand-searched. The risk of bias were used by Cochrane Collaboration's tool and the data were extracted. Meta-analysis was done with Revman 5.1. Nine articles met the eligibility criteria and were included, seven randomized controlled trials and 2 quasi-randomized controlled trials. Seven of these studies had unclear risk of bias and 2 had high risk of bias. Drainage could significantly increase 4.44 mm of the post-operative maximal mouth opening (P = 0.003), relief facial swelling (P < 0.05) and reduce post-operative complications (P = 0.008). But no evidence showed that drainage had a positive effects on post-operative pain (P = 0.09). Drainage could probably control the complications following impacted mandibular third molar extraction; but more randomized controlled trials are needed to reinforce the conclusion.


Regeneration and Repair of Skin Wounds: Various Strategies for Treatment.
Skin as a mechanical barrier between the inner and outer environment of our body protects us against infection and electrolyte loss. This organ consists of 3 layers: the epidermis, dermis, and hypodermis. Any disruption in the integrity of skin leads to the formation of wounds, which are divided into 2 main categories: acute wounds and chronic wounds. Generally, acute wounds heal relatively faster. In contrast to acute wounds, closure of chronic wounds is delayed by 3 months after the initial insult. Treatment of chronic wounds has been one of the most challenging issues in the field of regenerative medicine, promoting scientists to develop various therapeutic strategies for a fast, qualified, and most cost-effective treatment modality. Here, we reviewed more recent approaches, including the development of stem cell therapy, tissue-engineered skin substitutes, and skin equivalents, for the healing of complex wounds.


Trachoma control in two Central Australian Aboriginal communities: a case study.
This prospective case study assessed the additional impact of environmental changes (E) within the SAFE strategy in controlling trachoma in two Aboriginal communities (populations 315 and 385) in Central Australia. Baseline levels for trachoma, facial cleanliness, and nasal discharge were measured in children <15 years old. Health and facial cleanliness promotion were initiated in each community and housing and environmental improvements were made in one community. Azithromycin was distributed to all members of each community (coverage 55-73%). Assessments of trachoma and facial cleanliness were made at 3, 6, and 12 months post-intervention. Baseline trachoma rates were similar for the two communities (48 and 50%). Rates were significantly lower at 3, 6, and 12 months compared to baseline, but there was no significant difference between the two communities. The A/F components of the SAFE strategy significantly reduced the prevalence of trachoma; however, while the E intervention did not bring any apparent benefits, several factors might have masked them.


Long-term cryopreservation of peripheral blood stem cells in patients with advanced germ cell tumors using the dump-freezing method at -80 degrees C.
The objectives of this study were to evaluate the hematopoietic activity of peripheral blood stem cells (PBSCs) harvested from patients with advanced germ cell tumors after long-term cryopreservation in a mechanical freezer at -80 degrees C. Forty-one patients with advanced tumors were enrolled in this study. Flow cytometric analysis was performed to determine the total cell viability and the viability of CD34 positive cells of PBSCs stored between January 1995 and December 2001. The 41 patients were divided into two groups according to the duration of cryopreservation. In the 21 patients in whom the duration of storage was greater than 5 years, the total cell number did not change significantly before and after long-term cryopreservation (p=0.75). The percentage of viable cells and the total number of viable cells was significantly decreased after long-term cryopreservation (p<0.001 and p<0.05, respectively). However, the percentage of CD34 positive cells was significantly increased to 2.34% from 1.85% (p<0.05), resulting in non-statistically significant change in the total viable CD34 positive cells (p=0.98). A similar relation was found in the 20 patients whose storage duration was less than 2 years, although a significant change was found only in the percentage of viable cells. Moreover, pre-PBSCH factors, including age, cumulative dosage of cisplatin and the International Germ Cell Cancer Collaborative Group classification at the diagnosis did not affect the viable CD34 positive cell count after long-term cryopreservation. In conclusion, the present results suggest that CD34 positive cells could be successfully cryopreserved in a mechanical freezer at -80 degrees C for more than 5 years with no relation to pre-PBSCH factors and efficacy of PBSCH itself.


Which aspects of an oral health-related quality of life measure are mainly associated with global ratings of oral health in children?
To assess the associations of global subjective ratings of oral health with different aspects of an oral health-related quality of life measure, namely total Oral Impacts on Daily Performances (OIDP) and Child-Oral Impacts on Daily Performances (Child-OIDP) scores, extent and intensity of oral impacts, the different performance scores and oral impacts attributed to certain groups of oral conditions. This study was part of the Sixth Thailand National Oral Health Survey. One thousand and sixty-six 12-year-olds and 815 15-year-olds were interviewed on their OHRQoL using Child-OIDP and OIDP indices, respectively, and rated their overall oral problems on 3-point and 5-point scales, respectively. Associations of global rating of oral health with different Child-OIDP/OIDP outcome variables were estimated using Spearman's rank correlations and logistic regressions. Child-OIDP/OIDP scores were significantly associated with global ratings. Intensity was associated with global ratings, whereas extent was not. Impacts of 'very little' intensity were associated with a slightly higher chance of having poorer oral health perceptions. Global ratings were associated most with oral impacts attributed to dental caries, but not with the impacts related to natural process, such as eruption. Eating, cleaning teeth, emotional stability and smiling, plus social contact in 15-year-olds were performances associated with global ratings. Fifteen-year-old girls were more likely to perceive more oral problems than boys. Associations of global ratings of oral health with Child-OIDP/OIDP scores were mainly due to the intensity of impacts. Global ratings were related to multidimensional aspects of children's quality of life.


System development and integration in healthcare.
Nurse executives are challenged with integrating services within newly formed healthcare systems. The author provides a theoretical framework for system integration. The developmental process of integration in early system development, including the barriers to integration and methods to facilitate the process, are discussed.


Identification and quantification of protein S-nitrosation by nitrite in the mouse heart during ischemia.
Nitrate (NO3-) and nitrite (NO2-) are known to be cardioprotective and to alter energy metabolism in vivo NO3- action results from its conversion to NO2- by salivary bacteria, but the mechanism(s) by which NO2- affects metabolism remains obscure. NO2- may act by S-nitrosating protein thiols, thereby altering protein activity. But how this occurs, and the functional importance of S-nitrosation sites across the mammalian proteome, remain largely uncharacterized. Here we analyzed protein thiols within mouse hearts in vivo using quantitative proteomics to determine S-nitrosation site occupancy. We extended the thiol-redox proteomic technique, isotope-coded affinity tag labeling, to quantify the extent of NO2--dependent S-nitrosation of proteins thiols in vivo Using this approach, called SNOxICAT (S-nitrosothiol redox isotope-coded affinity tag), we found that exposure to NO2- under normoxic conditions or exposure to ischemia alone results in minimal S-nitrosation of protein thiols. However, exposure to NO2- in conjunction with ischemia led to extensive S-nitrosation of protein thiols across all cellular compartments. Several mitochondrial protein thiols exposed to the mitochondrial matrix were selectively S-nitrosated under these conditions, potentially contributing to the beneficial effects of NO2- on mitochondrial metabolism. The permeability of the mitochondrial inner membrane to HNO2, but not to NO2-, combined with the lack of S-nitrosation during anoxia alone or by NO2- during normoxia places constraints on how S-nitrosation occurs in vivo and on its mechanisms of cardioprotection and modulation of energy metabolism. Quantifying S-nitrosated protein thiols now allows determination of modified cysteines across the proteome and identification of those most likely responsible for the functional consequences of NO2- exposure.


HOXB9 inhibits proliferation in gastric carcinoma cells via suppression of phosphorylated-Akt and NF-κB-dependent Snail expression.
HOXB9 is a homeobox transcription factor which plays an important role in carcinoma development. This protein has been shown to inhibit cancer cell proliferation. However, the mechanisms that underpin HOXB9-mediated inhibition of cellular proliferation remain to be elucidated. In this study, two gastric cancer cell lines, SGC7901 and MKN45, were transfected with plasmids pLVX-HOXB9 and shHOXB9. These transfections resulted in the over-expression of the HOXB9 gene in the SGC7901/HOXB9 cells and knockdown of the HOXB9 gene in the MKN45/shHOXB9 cells. Over-expression of the HOXB9 gene in the SGC7901/HOXB9 cells caused an increase in the apoptotic rate and a concomitant reduction in metastatic ability compared with the knocked-down MKN45/shHOXB9 cells. Moreover, a reduction in the expression of the phosphorylated-Akt protein was observed in the SGC7901/HOXB9 cells, while an increase in expression of the same protein was observed in the MKN45/shHOXB9 cells. We also observed that HOXB9 mediated a reduction in both NF-κB and N-cadherin and Snail protein expression. Conversely, HOXB9 caused an increase in the expression of E-cadherin. In summary, this study reports that HOXB9 can suppress both phosphorylated-Akt expression and NF-κB activity. The latter phenomenon affects Snail protein expression and the inhibition of gastric carcinoma proliferation.


New aspects in molecular diagnosis of Lynch syndrome (HNPCC).
Colorectal carcinoma is one of the most common cancers that occurs in the human population, resulting in a mortality rate of more than 50%. Mismatch repair (MMR) defects are mostly manifested as high levels of microsatellite instability (MSI-H); this occurs in approximately 19% of all colorectal cancers including a smaller, but high-risk subgroup that is represented by hereditary nonpolyposis colorectal cancer (HNPCC, Lynch syndrome). The aim of molecular diagnosis is to distinguish these familial cases from of the majority of sporadic unstable cancers. Here, we review the widespread MSI-H phenotype in various tumours, assessing the genetic, epigenetic and morphological differences to distinguish between sporadic and familial unstable colorectal cancers. The most important morphological feature is the presence of tumour-infiltrating lymphocytes (TILs), the presence of which is typical in MSI cancers. The main discriminators for HNPCC patients are family history, age at onset up to 60 years and MMR defects caused by germline mutations. This is in contrast to the discriminators for sporadic colorectal cancers, which include onset at any age, BRAF mutation presence and epigenetically MMR inactivation by extensive CpG-island methylation. This article presents a modified strategy for molecular diagnosis of HNPCC by selective inclusion of recently recognized characteristics of tumours. The clear identification of affected families can improve the strategy of early detection, therapy and prevention of colorectal cancers.


Outcomes of COVID-19 Omicron variant in patients with rheumatoid arthritis: a nationwide Greek cohort study.
Patients with rheumatoid arthritis (RA) were at increased risk for COVID-19-associated hospitalization and death during the first year of pandemic in Greece. We aimed to examine their outcomes after SARS-Cov-2 Omicron, a more contagious but with milder clinical impacts variant prevailed. A retrospective, nationwide study was conducted between 1-January-2022 and 30-June-2022 in all RA patients under treatment and matched (1:5) on age, sex, and region of domicile random general population comparators. Confirmed SARS-CoV-2 infections, hospitalizations and deaths, anti-rheumatic medications, prior COVID-19, vaccinations and anti-viral medications were recorded. Among 34 182 RA patients, infections (n = 5,569, 16.29%), hospitalizations (n = 489, 1.43%) and deaths (n = 106, 0.31%) were more frequent than comparators. Incidence rates per 1000 person/years of infection [IRR (95%CI):1.19 (1.16-1.23)], hospitalization [IRR (95%CI):2.0 (1.82-2.24)], and death [IRR (95%CI):1.81 (1.44-2.27)] were increased in RA despite better vaccination coverage (89% vs 84%) and more frequent use of anti-viral medications (2.37% vs 1.08). Logistic regression analysis after correcting for age, sex, vaccinations, prior COVID-19, anti-viral and anti-rheumatic medications in SARS-CoV-2 infected RA patients and comparators revealed increased risk of hospitalization (OR : 2.02, 95%CI : 1.79- 2.27) and death [OR : 1.73, (95%CI : 1.36-2.20)] in RA. Among infected RA patients rituximab treatment conferred increased risks for hospitalization [OR: 6.12, (95%CI : 2.89-12.92)] and death [OR : 12.06 (95%CI : 3.90-37.31)], while JAK inhibitors increased only hospitalization risk [OR : 2.18 (95%CI : 1.56-3.06)]. RA remains a risk factor for hospitalization and death in an era of a relatively low COVID-19 fatality rate, pointing to the need of perseverance in vaccination programs and wider use of anti-viral medications.


Cost assessment of treatment of acute myocardial infarction and angiographically visible coronary thrombus.
Study was aimed to assess the real-world costs of manual thrombectomy (MT) in selected ST-segment elevation myocardial infarction patients with intracoronary thrombus (IT). Study group (IT+) comprised 51 patients with MT applied and control group (IT-) comprised 56 patients without IT who underwent angioplasty alone. Costs comprised hospital care and cost of disposable materials used during primary angioplasty. Complex management of patients with IT is more expensive, though allows to achieve clinical outcomes comparable to low-risk ST-segment elevation myocardial infarction patients without IT. A complex pharmaco-interventional strategy, with glycoprotein IIB/IIIA inhibitor and MT, though more expensive, may prove cost-effective.


Inhibition of FAK and VEGFR-3 binding decreases tumorigenicity in neuroblastoma.
Neuroblastoma is the most common extracranial solid tumor of childhood and is responsible for over 15% of pediatric cancer deaths. Focal adhesion kinase (FAK) is a nonreceptor tyrosine kinase that is important in many facets of tumor development and progression. Vascular endothelial growth factor receptor-3 (VEGFR-3), another tyrosine kinase, has also been found to be important in the development of many human tumors including neuroblastoma. Recent reports have found that FAK and VEGFR-3 interact, and we have previously shown that both of these kinases interact in neuroblastoma. We have hypothesized that interruption of the FAK-VEGFR-3 interaction would lead to decreased neuroblastoma cell survival. In the current study, we examined the effects of a small molecule, chloropyramine hydrochloride (C4), designed to disrupt the FAK-VEGFR-3 interaction, upon cellular attachment, migration, and survival in two human neuroblastoma cell lines. We also utilized a murine xenograft model to study the impact of C4 upon tumor growth. In these studies, we showed that disruption of the FAK-VEGFR-3 interaction led to decreased cellular attachment, migration, and survival in vitro. In addition, treatment of murine xenografts with chloropyramine hydrochloride decreased neuroblastoma xenograft growth. Further, this molecule acted synergistically with standard chemotherapy to further decrease neuroblastoma xenograft growth. The findings from this current study help to further our understanding of the regulation of neuroblastoma tumorigenesis, and may provide novel therapeutic strategies and targets for neuroblastoma and other solid tumors of childhood.


Antibacterial activity and adhesive properties of a chitosan-containing dental adhesive.
To evaluate the antibacterial activity of adhesive resin incorporating chitosan as well as the adhesive characteristics. An experimental adhesive was prepared by adding 0.12%, 0.25%, 0.5%, and 1% (w/w) chitosan solution to Single Bond adhesive resin. The solution of chitosan was prepared by dissolving 2 g of chitosan powder in 1 liter of 1% (v/v) acetic acid. Single Bond without chitosan was used as a control. The antibacterial activity was evaluated using a direct contact test against Streptococcus mutans. The viscosity, degree of conversion, pH, and microtensile bond strength (ΜTBS) values of the experimental adhesives to dentin were evaluated. Data were analyzed using the ANOVA and Tukey tests. Statistical significance was set at the .05 probability level. The antibacterial properties of freshly prepared and aged experimental adhesives incorporating chitosan were found to exhibit an inhibitory effect on the growth of Streptococcus mutans compared with the unmodified adhesive resin (P < .05). The viscosity of the experimental adhesives increased with increasing the concentrations of chitosan incorporation into the adhesive. However, the degree of conversion and pH values decreased with increasing the concentrations of chitosan incorporation into the adhesive. Experimental adhesives incorporating 0.12% and 0.25% (w/w) chitosan showed no significant differences in the ΜTBS values compared with the control (P > .05). However, the incorporation of 0.5% and 1% (w/w) chitosan into the dental adhesive significantly decreased the ΜTBS (P < .05). Adhesive resin that contains 0.12% (w/w) chitosan is a promising antibacterial adhesive that does not adversely affect adhesive properties.


Clinical Outcomes of Total Hip Arthroplasty With the Anterolateral Modified Watson-Jones Approach for Displaced Femoral Neck Fractures.
In recent years, many studies have reported good results with total hip arthroplasty (THA) for displaced femoral neck fractures (FNFs). However, no study has reported the clinical outcomes of the anterolateral modified Watson-Jones THA (MWJ-THA) for displaced FNFs. This study aimed to investigate the clinical results of THA for displaced FNFs at our hospital and to discuss the advantages of MWJ-THA over THA with other approaches for displaced FNFs. Forty-three patients who underwent MWJ-THA for displaced FNFs were included in this study. Patient characteristics, preinjury walking ability, activities of daily living, implants used, walking ability (at 1, 3, and 6 months after surgery), cup placement angle, clinical hip score, surgical complications, revision surgery, and death within 1 year after surgery were investigated. The mean age of the 43 patients was 63.3 years, and the mean body mass index (kg/m2) was 21.1. Regarding the heads used, 28-mm heads were used in 4 patients, 32-mm heads were used in 32 patients, and 36-mm heads were used in 7 patients. The cups were placed in the Lewinnek safety zone (93.0%). Four patients had stem sinkage of a few millimeters. 6 months postoperatively, 38 patients walked unaided, and 4 patients walked with a cane. The Harris Hip Score averaged over 90 points at all time points. No postoperative dislocation was observed. Two patients died within 1 year postoperatively. In this study, MWJ-THA was performed for displaced FNFs and resulted in no postoperative dislocations. Furthermore, more than 90% of the patients regained their preinjury walking ability at 6 months postoperatively. MWJ-THA has great dislocation control and is effective in treating displaced FNFs.


Long-term transgene expression in mouse neural progenitor cells modified with phiC31 integrase.
Stem cells can potentially be utilized in combined gene/cell therapies for neural diseases. We examined the ability of the non-viral phiC31 integrase system to promote stable transgene expression in mouse neural progenitor cells (mNPCs). phiC31 integrase catalyzes the sequence-specific integration of attB-containing plasmids into pseudo attP sites in mammalian genomes, to produce long-term transgene expression. We achieved gene transfer by co-nucleofection of a plasmid carrying the luciferase marker gene and an attB site and a plasmid expressing integrase in mNPCs that had been generated in a neurosphere preparation. Luciferase expression was quantified in live cells for 8 weeks, revealing persistence of gene expression. Sequence-specific integration at a preferred pseudo attP site in the mouse genome was detected by using PCR. Furthermore, sustained transgene expression was demonstrated in genetically modified NPCs that were cultured in conditions that promoted either growth or differentiation into neurons and astrocytes. Our results demonstrate that the phiC31 integrase system produces stable transgene expression in adult mNPCs and their progeny and may be useful in strategies for combating neurodegenerative disorders.


The enhancing mechanism of capric acid (C10) from a suppository on rectal drug absorption through a paracellular pathway.
Capric acid (C10) enhanced the absorption of cefoxitin sodium in a concentration-dependent manner following the rectal administration as a suppository in rats. The optimal concentration of C10 was 13%. C10 administered as a suppository also reduced rectal membrane resistance (Rm), showing that the above enhancing effect was induced by widening the paracellular pathway. Both the enhancing effect on the absorption and the reducing effect on Rm were inhibited by W7, an inhibitor of myosin light chain kinase. These results supported that, as shown in the in vitro Caco-2 cell system, the C10 effect on the paracellular pathway is due to activating the contraction of Ca(2+)-calmodulin-dependent actin filament.


Effects of a 10% carbamide peroxide bleaching agent on roughness and microhardness of packable composite resins.
Bleaching agents containing 10% carbamide peroxide may be applied to the surface of preexisting packable resin-based composite restorations. The aim of this in vitro study was to evaluate the effect of a 10% carbamide peroxide bleaching agent (Review, SS White, Rio de Janeiro, Brazil) on surface roughness and microhardness of three packable resin-based composites (Fill Magic condensable, Vigodent, Rio de Janeiro, Brazil; Alert, Jeneric Pentron, Wallingford, CT, USA; Definite, Degussa, Hanau, Germany). For the control (no bleaching) and experimental (bleaching treatment) groups, 12 specimens of each material were prepared in cylindrical acrylic molds. The experimental specimens were exposed to the bleaching agent for 6 hours a day for 3 weeks. During the remaining time (18 h), they were stored in artificial saliva. The control specimens remained immersed in artificial saliva throughout the experiment. Surface roughness and microhardness measurements were performed on the top surface of each specimen. Analysis of variance and the Tukey test showed no significant differences in roughness among the packable composites evaluated (p = .18), but those submitted to the treatment with a 10% carbamide peroxide bleaching agent displayed significantly higher mean surface roughness than did the corresponding control group for each material. For the microhardness tests, there were significant differences among materials (p < .0001). Alert showed the highest microhardness values followed by Definite and Fill Magic condensable. Ten percent carbamide peroxide bleaching agents may change the surface roughness of packable composites, but they do not alter their microhardness. Under the conditions of this study, a 3-week bleaching regimen with 10% carbamide peroxide agents affected the roughness of packable composites. Nevertheless, the surface microhardness was not affected, although there were differences among the materials evaluated inherent to their composition.


[Efficacy of polymeric 239Pu removal by liposome-encapsulated pentacin].
Liposome-incapsulated pentacine (DTPA) removes intracellular polimeric 239Pu and colloidal hydroxide of polymeric 239Pu from a rat body in the amounts 2- and 4 times, respectively, exceeding those eliminated by free DTPA. However the amount of 239Pu removed decreases sharply with increasing 239Pu hydrolysation and polymerization. It is concluded that the effectiveness of 239Pu removal depends on the physico-chemical status of the radionuclide and its interaction with the biosubstrate rather than on the amount of DTPA entering the cells.


Policy Effects of Ecological Red Lines on Industrial Upgrading and Health Promotion: Evidence From China Based on DID Model.
The implementation of the Ecological Red Lines (ERL) policy in China is under the background that natural resources have been immoderately exploited for serving rapid economic growth in the last 40 years, where the ecosystem's degradation happened and people's health could be affected. As the secondary industry is the contribution source of rapid growth as well as the threat source that threatens the natural environment and public health, the delimitation of ERL can act as a legal restriction that forces the industries to control the emissions and to upgrade the industrial composition. This paper conducts an ex-post policy evaluation on the improvement effects of industrial structure and residents' health and through ERL's pilot scheme in four provinces of China. By using the difference-in-differences (DID) method, the estimation results show that: (1) The industrial upgrading effect exists but to a small extent, as the ERL policy has generally elevated the tertiary industry's output by only 0.033% and hardly shown any promotion effects on the ratio of the tertiary industry to secondary industry; (2) The residents' health has been significantly improved by 1.029% after ERL policy on the whole, and enhanced over time mostly; (3) The health promotion effects are similar among three out of the four pilot provinces, whereas the industrial upgrading effects performed large heterogeneities among the four. These empirical results may provide references for the wider extension of ERL policy with more practical execution solutions in developing economies.


Characterization of TistH, a multifunctional peptide from the scorpion Tityus stigmurus: Structure, cytotoxicity and antimicrobial activity.
The presence of bioactive peptides in animal venoms has been targeted in scientific research for assessing biological activities, as well as mechanisms of action. A recent study by our group observed hypotensive action of TistH (Tityus stigmurus Hypotensin), a peptide deduced from the transcriptome of T. stigmurus venom gland. The present study aims to analyze TistH structure properties and to evaluate its toxicity on normal and tumor cells, its in vitro antimicrobial activity, as well as its inflammatory effect. Circular dichroism analyses of TistH showed a general predominance of α-helix conformation in TFE (20-70%) and structural stability to pH variations. TistH was not cytotoxic to normal cell lines (3T3, RAW and HEK), and also not to cancer cell lines (HeLa, B16, 786-0, SiHa and HepG2). The peptide did not present inflammatory activity up to 6 h after administered subcutaneously to Swiss mice. It was observed that concentrations of 4-1024 μg/mL of TistH produced no inhibition against the bacteria Staphylococcus aureus, Staphylococcus epidermidis e Pseudomonas aeruginosa. The results of antifungal assays showed a moderate activity of TistH against Candida albicans strain LM-108 and the filamentous fungus Trichophyton rubrum LM-640, with growth inhibition at a concentration of 1024 μg/mL. In contrast, the peptide presented a greater activity (MIC 128 μg/mL) against C. albicans LM-106, Candida tropicalis ATCC 13308 and Aspergillus flavus strains LM-247 and LM-26, fungi that cause oral and vaginal infections, candidiasis and respiratory allergies, respectively. The present data contribute to a better understanding of TistH and its possible use as a bioactive compound. This multifunctional peptide is capable of acting as anti-hypertensive, as well as to inhibit the growth of fungal strains, having low toxicity, which suggests its safety for using as a pharmacological agent.


Impaired functional capacity of fetal endothelial cells in preeclampsia.
Preeclampsia is one of the main contributers to maternal and fetal morbidity and mortality during pregnancy. A history of preeclampsia puts mother and offspring at an increased cardiovascular risk in later life. We hypothesized that at the time of birth functional impairments of fetal endothelial cells can be detected in pregnancies complicated by preeclampsia and that a therapeutic intervention using 1,25 (OH)2 vitamin D3 can reverse the adverse effects of preeclampsia on cell function. Human umbilical vein endothelial cells (HUVEC) were isolated from umbilical cords obtained from preeclamptic (N = 12) and uncomplicated pregnancies (N = 13, control). Placental villous tissue fragments from uncomplicated term pregnancies were incubated in explant culture for 48 h at 2% (hypoxia), 8% or 21% O2. Explant conditioned media (CM) was collected and pooled according to oxygen level. We compared the ability of preeclampsia vs. control HUVEC to migrate, proliferate, and form tubule-like networks in a Matrigel assay, in the presence/absence of CM and 1,25(OH)2 vitamin D3. HUVEC from preeclamptic pregnancies showed reduced migration (P = 0.04) and tubule formation (P = 0.04), but no change in proliferation (P = 0.16) compared to healthy pregnancies. Placental villous explant CM derived from 2% O2 incubations significantly reduced HUVEC migration, when compared to non-CM (P = 0.04). Vitamin D3 improved HUVEC function in neither of the groups. There was no significant difference in VEGF gene expression between healthy and preeclamptic pregnancies and no effect of Vitamin D3 on VEGF expression. Reduced functional abilities of fetal endothelial cells from preeclamptic pregnancies suggests that disease pathways, possibly originating from the dysfunctional placenta, negatively impact fetal endothelium. The neutral effect of 1,25(OH)2 vitamin D3 contrasts with previous findings that vitamin D rescues the poor migration, proliferation and tubule formation exhibited by cord blood fetal endothelial progenitor cells from preeclamptic pregnancies. Further investigations to distinguish pathways by which offspring exposed to preeclampsia are at risk for cardiovascular disease are needed.


Immune reconstitution inflammatory syndrome manifesting as disseminated tuberculosis, deep venous thrombosis, encephalopathy and myelopathy.
We describe an unusual case of a 25-year-old human immunodeficiency virus (HIV)-positive male with a pre-treatment CD4 count of 144 cells/microL, who had received highly active antiretroviral therapy (HAART) consisting of lamivudine, stavudine and nevirapine for three months, developing immune reconstitution inflammatory syndrome (IRIS) manifesting as disseminated tuberculosis (TB), myelopathy, encephalopathy and deep venous thrombosis (DVT). In addition to HAART and antituberculosis treatment (ATT), the patient was given non-steroidal anti-inflammatory drugs, oral vitamin B12 and heparin, which was later switched to oral warfarin.


Tc99m-MDP uptake in ascitic fluid in a patient with prostate carcinoma: A clue to detect metastases.
Bone scintigraphy with Tc-99m methylene diphosphonate (MDP) is used to detect metastases in patients with cancer. Uptake in non-osseous, non-urologic tissues is occasionally found in the routine bone scintigraphy, which may mimic as metastatic lesions. The authors describe the case of a 70-year-old man with prostate cancer, showing diffuse tracer uptake in the left hemithorax and entire abdomen on bone scan that required additional imaging modality for localization. Careful interpretation is needed of the unusual uptake of radiotracer in regions other than the skeleton for metastatic work up.