Induction patterns of new CYP1 genes in environmentally exposed rainbow trout.
The cytochrome P4501 (CYP1) gene family comprises four subfamilies in fish: CYP1A, CYP1B, CYP1C, and CYP1D. Only two CYP1 genes, CYP1A1 and CYP1A3, are so far known in rainbow trout (Oncorhynchus mykiss). The present study aimed to identify other CYP1 subfamily genes in rainbow trout, to establish methods for quantitative mRNA expression analysis of these genes, and to determine their basal and induced mRNA expression in gills and liver. Another goal was to examine their mRNA expression in environmentally exposed fish. We cloned four new transcripts, denoted rbCYP1B1, rbCYP1C1, rbCYP1C2, and rbCYP1C3. Levels of these and the previously known rbCYP1A transcripts were determined by real-time PCR in unexposed fish, fish exposed to the potent aryl hydrocarbon receptor (AhR) agonist 3,3',4,4',5-pentachlorobiphenyl (PCB126), and fish caged in various waters in the Uppsala region (Sweden). The mRNA expression patterns observed in unexposed rainbow trout (basal levels) were markedly similar to those reported for orthologous genes in other species. All six transcripts were induced by PCB126 in gills and liver, suggesting all genes to be AhR regulated. The caged fish showed clear rbCYP1 induction in gills at all monitoring sites (up to 70-fold the basal level), whereas the liver responses were weak; induction (up to 5-fold) was recorded only at the Uppsala municipal sewage treatment plant outlet. Gill filament EROD activity was induced at all caging sites. Most interestingly, the rbCYP1 gene response patterns in gills differed among caging sites and among subfamilies. The EROD induction seemed to only reflect induction of rbCYP1A transcription. Response patterns of multiple CYP1 genes in gills and liver could provide an improved monitoring strategy. Such patterns could be used to characterize complex mixtures of AhR agonists and antagonists in aquatic environments.


Prolonged high-dose intravenous magnesium therapy for severe tetanus in the intensive care unit: a case series.
Tetanus rarely occurs in developed countries, but it can result in fatal complications including respiratory failure due to generalized muscle spasms. Magnesium infusion has been used to treat spasticity in tetanus, and its effectiveness is supported by several case reports and a recent randomized controlled trial. Three Caucasian Greek men aged 30, 50 and 77 years old were diagnosed with tetanus and admitted to a general 12-bed intensive care unit in 2006 and 2007 for respiratory failure due to generalized spasticity. Intensive care unit treatment included antibiotics, hydration, enteral nutrition, early tracheostomy and mechanical ventilation. Intravenous magnesium therapy controlled spasticity without the need for additional muscle relaxants. Their medications were continued for up to 26 days, and adjusted as needed to control spasticity. Plasma magnesium levels, which were measured twice a day, remained in the 3 to 4.5 mmol/L range. We did not observe hemodynamic instability, arrhythmias or other complications related to magnesium therapy in these patients. All patients improved, came off mechanical ventilation, and were discharged from the intensive care unit in a stable condition. In comparison with previous reports, our case series contributes the following meaningful additional information: intravenous magnesium therapy was used on patients already requiring mechanical ventilation and remained effective for up to 26 days (significantly longer than in previous reports) without significant toxicity in two patients. The overall outcome was good in all our patients. However, the optimal dose, optimal duration and maximum safe duration of intravenous magnesium therapy are unknown. Therefore, until more data on the safety and efficacy of magnesium therapy are available, its use should be limited to carefully selected tetanus cases.


Cross-reactivity of monoclonal antibodies to defined human leucocyte differentiation antigens with bovine cells.
Thirty-seven subpanels of monoclonal antibodies (mAbs) included within the Vth International Workshop on Human Leucocyte Differentiation Antigens (Vth Workshop) were assayed for reactivity with bovine peripheral blood leucocytes. Sixty-five of the 772 mAbs (8.4%) stained bovine cells. mAbs from each of the 27 different CD groups that contained a mAb reacting with cattle were further investigated to compare the cellular expression of the antigen in cattle with that reported for the different CD antigens in humans. Two-colour immunofluorescence staining of the Vth Workshop mAbs against characterized bovine leucocyte subpopulation markers that identified monocytes, B cells, CD4, CD8 and WC1 +T cells were used for these analyses. Eighteen of the mAbs to different human CD antigens (CD11a, CD14, CD18, CD21, CD27, CD29, CD49a, CD49b, CD49d, CD49e, CD51, CD61, CD62L, CD62P, CD63, CDw78, CD98, CD100) stained bovine antigens with an almost identical cellular distribution to that reported in humans. This implies that these mAb react with the homologous cattle molecules. Nine mAbs (CD35, CD37, CD49c, CD50, CD54, CD66, CD81, CD88, CD102) stained bovine cells but the cellular distribution of the bovine antigen was different to that reported in humans implying either a different cellular distribution for these antigens in cattle or a reaction with a different molecule. The investigation has allowed the identification of several bovine homologues of human CD antigens that have not been previously defined in cattle and the cross-reacting mAbs will be valuable reagents for future investigations of bovine immunology.


AN ULTRAMICROSCOPIC STUDY OF THE TWO STAGES OF BLOOD COAGULATION.
There is no reaction-basis for the division of the process of blood-coagulation into the two stages (transparent and opaque) which appearso strikingly in sloiwly-clotting mammalian blood. These stages are saperficial phenomena which merely reflect the extent of fibrin-formation. Coagulation is a gradual continuous process of fibrin-formation; and in the clotting of normal plasma, fibrin needles can be demonstrated in the earliest appreciable coagulum, however delicate, transparent or gellike.


Coordination behavior of bis-phenolate saturated and unsaturated N-heterocyclic carbene ligands to zirconium: reactivity and activity in the copolymerization of cyclohexene oxide with CO2.
Tetravalent zirconium complexes supported by tridentate bis-phenolate imidazolidin-2-ylidene (L1), imidazol-2-ylidene (L2) and benzimidazol-2-ylidene (L3) NHC ligands were synthesized and evaluated as precursors for the copolymerization of cyclohexene oxide (CHO) with CO2. While the reactivity of the imidazolidinium [H3L1] chloride salt with Zr(OiPr)4(HOiPr), and subsequent ligand exchanges with either (CH3)3SiCl or LiOiPr lead to a series of heteroleptic compounds (κ3-O,C,O-L1)Zr(X)2(THF) (X = Cl, OiPr), both imidazolium [H3L2] and benzimidazolium [H3L3] chloride salts give a mixture of homoleptic (κ3-O,C,O-NHC)2Zr and zwitterionic (κ2-O,O-HL)ZrCl2(OiPr) compounds along with traces or the absence of the heteroleptic intermediate (κ3-O,C,O-NHC)Zr(Cl)(OiPr)(THF). Such dissimilar reactivity between the unsaturated and saturated NHC ligands is predominantly ascribed to the increased acidity of azolium salts along with the π-donor strength of the Ccarbene in L2 and L3-Zr moieties. The reactivity with the more acidic azolium salts (H3L2/3) and the destabilized Zr-Xtrans to NHCcarbene bond results in a significant increase in the amount of homoleptic compounds generating HCl. The released HCl reacts preferentially with the heteroleptic intermediates having non-planar NHC ligands (i.e. L2/3) promoting the formation of zwitterionic complexes. The in situ deprotonation of the isolated zwitterionic (κ2-O,O-HL3)ZrCl2(OiPr) compound by using Ag2O gives the homoleptic complex as the major component along with a bimetallic hydroxo-bridged [(κ3-O,C,O-L3)Zr(μ-OH)(OiPr)]2 compound. Of particular interest is that only the heteroleptic NHC-Zr(iv) complexes were identified to be active and highly selective towards the copolymerization of CHO with CO2 independently of the co-catalysts used (both anionic and neutral) under mild conditions (PCO2 < 1 bar, T = 60 °C), and gave atactic and completely alternating copolymers in a controlled manner (Mw/Mn ≈ 1.3-1.8). In contrast, the isolated homoleptic, zwitterionic and bimetallic zirconium species were found to be inactive under similar reaction conditions. Although the activity found for NHC-Zr(iv) complexes is nearly of the same order of magnitude as that of the NHC-Ti(iv) analogues, these results are the first examples of tetravalent zirconium complexes achieving high selectivity (99% in PCHC) in the catalyzed copolymerization of CHO with CO2.


Neuropsychological features of mild cognitive impairment and preclinical Alzheimer's disease.
Recent research has identified a transitional state between the cognitive changes of normal aging and Alzheimer's disease (AD), known as mild cognitive impairment (MCI). MCI patients experience memory loss to a greater extent than one would expect for age, yet they do not meet currently accepted criteria for clinically probable AD. An issue currently under investigation is whether MCI represents the preclinical stages of AD or a distinct and static cognitive aetiology. In an attempt to address this issue, the present investigations are adopting a convergent approach to the detection of preclinical AD, where multiple risk factors are considered when making a diagnosis. Currently, one of the most important tools when assessing early cognitive changes is neuropsychological evaluation. MCI subjects typically record neuropsychological performance between that of healthy older individuals and demented patients. Tests assessing new learning, delayed recall and attention/executive function seem to provide valuable information for screening and diagnosis of MCI and early AD if interpreted properly. Recommendations concerning methodological issues and the early management of neuropsychological MCI studies were made.


Cell determination and differentiation of identified serotonin-immunoreactive neurons in the grasshopper embryo.
We have begun to investigate the factors that underlie neurotransmitter determination in the central nervous system of the grasshopper embryo. The most prominent serotonin-immunoreactive neurons in the segmental ganglia are three clonally related interneurons, cells S1, S2, and S3. S1 and S2 are sibling neurons and are the first two born in the family of neurons that is produced by neuroblast 7-3; cell S3 derives from one of the second-born pair in the same family. S1 is serotonin-immunoreactive in all thoracic and abdominal segments, S2 in all but two segments (T3 and A1), and S3 in only the prothorax (T1). These segment-specific differences are not due to differential cell death but rather can be ascribed to biochemical differences between lineally homologous neurons. Furthermore, these homologous neurons also display interesting segment-specific differences in their morphology. Laser ablation of neuroblast 7-3 before it begins its series of programmed cell divisions results in the absence of its normal serotonin-immunoreactive neuronal progeny when screened at mature stages. These experiments support the hypothesis that transmitter determination is at least in part regulated by cell lineage in the grasshopper embryo and that some clonally related neurons appear to share a common transmitter determination by virtue of their common lineage. The results further suggest that cell lineage regulation of transmitter determination may vary according to segment or may be modified by as yet unidentified segment-specific factors.


Bisphosphonate-related osteonecrosis of the jaw: is pH the missing part in the pathogenesis puzzle?
Bisphosphonate-related osteonecrosis of the jaw (BRONJ) is a side effect of bisphosphonate therapy, primarily diagnosed in patients with cancer and metastatic bone disease and receiving intravenous administrations of nitrogen-containing bisphosphonates. If diagnosis or treatment is delayed, BRONJ can develop to a severe and devastating disease. Numerous studies have focused on BRONJ, with possible pathomechanisms identified to be oversuppression of bone turnover, ischemia due to antiangiogenetic effects, local infections, or soft tissue toxicity. However, the precise pathogenesis largely remains elusive and questions of paramount importance await to be answered, namely 1) Why is only the jaw bone affected? 2) Why and how do the derivatives differ in their potency to induce a BRONJ? and 3) Why and when is BRONJ manifested? The present perspective reflects on existing theories and introduces the hypothesis that local tissue acidosis in the jaw bone offers a conclusive pathogenesis model and may prove to be the missing link in BRONJ.


PAX3 and PAX7 as upstream regulators of myogenesis.
Like other subclasses within the PAX transcription factor family, PAX3 and PAX7 play important roles in the emergence of a number of different tissues during development. PAX3 regulates neural crest and, together with its orthologue PAX7, is also expressed in parts of the central nervous system. In this chapter we will focus on their role in skeletal muscle. Both factors are key regulators of myogenesis where Pax3 plays a major role during early skeletal muscle formation in the embryo while Pax7 predominates during post-natal growth and muscle regeneration in the adult. We review the expression and functions of these factors in the myogenic context. We also discuss mechanistic aspects of PAX3/7 function and modulation of their activity by interaction with other proteins, as well as the post-transcriptional and transcriptional regulation of their expression.


In situ distinction between steroid receptor binding and transactivation at a target gene.
We have developed a DNA interference assay in the yeast Saccharomyces cerevisiae that is designed to indicate the intracellular DNA-binding status of the estrogen receptor. The assay utilizes a promoter containing multiple copies of a GAL4-estrogen receptor binding sequence. This element is designed so that either an estrogen receptor or a GAL4 molecule, but not both, can occupy it simultaneously. The assay is extremely sensitive, and at concentrations of estrogen receptor below that required for maximal transcriptional activation of its target estrogen response element, a quantitative inhibition of GAL4-mediated transcription is seen. Inhibition occurs thought the disruption of complex cooperative interactions among the GAL4 molecules in this reporter. The data obtained from our experiments show that at low concentrations of receptor, hormone is required to promote DNA binding. Overexpression of receptor leads to occupation of the estrogen receptor element in the absence of ligand. In contrast, this latter receptor form will not activate transcription. Our results are consistent with a two-step process for receptor activation. Ligand first causes dissociation of receptor from an inhibitory complex within the cell and produces a DNA-binding form. Second, it converts receptor to a transcriptionally competent form. With use of this yeast model system, these two steps can be distinguished in situ.


Treatment of atlantoaxial instability in pediatric patients.
The atlantoaxial region has been extensively described as a spinal segment especially prone to injury in children. In this clinical review, the authors evaluate and summarize the management of 23 pediatric cases of atlantoaxial instability treated between March 1990 and October 2002. Four broad categories of atlantoaxial problems were observed-atlantoaxial rotatory subluxation in six patients, anterior-posterior atlantoaxial instability caused by ligamentous injury or congenital ligamentous laxity (10 patients), atlantoaxial fracture with or without dislocation (five patients), and atlantooccipital dislocation (two patients). Most cases (60.9%) were treated without surgical intervention, resulting in excellent outcomes; however, 21.7% of cases were treated with a cervical halo (mean patient age 72.6 months) alone for 3 months. Various techniques of surgical stabilization including transarticular screws with sublaminar wiring, transoral decompression with posterior plating, and laminectomy with Steinmann pin occipital-cervical fusion were used with good results. Both patients with atlantooccipital dislocation underwent immediate Locksley occipital-cervical fusion, with marked neurological improvement. Individualized case management must be based on clinical presentation, with internal fixation being the last resort.


Prevalence and incidence of high risk for obstructive sleep apnea in World Trade Center-exposed rescue/recovery workers.
World Trade Center (WTC)-exposed rescue/recovery workers continue to have high rates of gastroesophageal reflux disease (GERD), chronic rhinosinusitis, and posttraumatic stress disorder (PTSD) symptoms. This study examines the relationship between these WTC-related conditions and being at high risk for obstructive sleep apnea (OSA). The Fire Department of the City of New York (FDNY) performs periodic health evaluations on FDNY members every 12 to 18 months. Evaluations consist of physician examinations and self-administered health questionnaires, which, since 2005, have incorporated questions about sleep problems that were adapted from the Berlin Questionnaire. The study population consisted of 11,701 male firefighters and emergency medical service personnel. Incidence analyses were limited to a cohort (n = 4,576) who did not meet the criterion for being at high risk for OSA at baseline (between September 12, 2005 and September 8, 2006) and had at least one follow-up assessment, on average, 1.4 (±0.5) years later. The baseline prevalence of high risk for OSA was 36.5%. By follow-up, 16.9% of those not at high risk initially became at high risk for OSA. In multivariable logistic regression models predicting incident high risk for OSA, independent predictors included: earlier time of arrival at the WTC site, GERD, chronic rhinosinusitis, PTSD symptoms, self-assessed fair/poor health, low body mass index (BMI < 18.5 kg/m(2)), and, as expected, BMI > 30 kg/m(2) and weight gain of ≥10 lb (4.5 kg). We found significant associations between being at high risk for OSA and common WTC-related conditions, although the responsible causative mechanisms remain unknown. Since the etiology of OSA is likely multifactorial, improvement may require successful treatment of both OSA and its comorbid conditions.


Coexistence in field samples of two variants of the infectious salmon anemia virus: a putative shift to pathogenicity.
Genetic reassortment plays an important role in the evolution of several segmented RNA viruses and in the epidemiology of their associated diseases. In particular, orthomyxoviruses show rapid fluctuation in the proportion of viral variants coexisting in an infected individual, especially under strong selective pressure. This is particularly relevant in salmon production carried out under confined and stressful conditions where one of the most feared pathogenic agents is the Infectious Salmon Anemia Virus, an orthomyxovirus family member whose biological behavior is only recently beginning to be understood. Pathogenicity of the virus has been mainly associated with deletions of the HPR region in coding segment 6 and the presence or absence of a specific insertion in a key region in coding segment 5. In this study we report, for the first time in Chile, the coexistence of two variants in fully asymptomatic fish. Of five samples analyzed, two were identified as the non-pathogenic variant, HPR0, and two as the highly pathogenic HPR7b variant, though with no clinical signs detectable in the fish. Interestingly, one of the samples unequivocally carried both variants, again without any clinical signs. Considering that in none of the samples the typical insertion in coding segment 5 was detected, it is our impression that this may represent a shift from the non-pathogenic HPR0 variant towards the highly infective HPR7b variant. If this were the case, the transition may be triggered first by deleting the corresponding sequence of the HPR region of segment 6, followed by the putative insertion in segment 5 to generate a virulent strain.


Exploring the Impact of Morphology on the Properties of Biodegradable Nanoparticles and Their Diffusion in Complex Biological Medium.
Nanoparticle morphology (size, shape, and composition) and surface chemistry are the determining factors underpinning the efficacy of such materials in therapeutic applications. The size, shape, and surface chemistry of a nanoparticle can strongly influence key properties such as interactions with diverse biological fluids and interfaces and, in turn, impact the delivery of bioactive cargo, modulating therapeutic performance. This is exemplified in ocular drug delivery, where potential therapeutics must navigate complex biological media such as the gel-like vitreal fluid and the retina. Biodegradable block copolymer amphiphiles are a robust tool for the engineering of various types of self-assembled nanoparticles with diverse morphologies ranging from spherical and tubular polymersomes to spherical and worm-like micelles. Here, we explore the effect of morphological features such as shape and surface chemistry upon the interactions of a series of copolymer nanoparticles with retinal (ARPE-19) cells and the release of a low solubility drug (dexamethasone) that is currently used in ocular therapy and study their diffusion in vitreous using ex vivo eyes. We demonstrate that both aspect ratio and surface chemistry of nanoparticles will influence their performance in terms of cell uptake, drug release, and diffusion with high aspect ratio shapes demonstrating enhanced properties in relation to their spherical counterparts.


Variation Among Pediatric Orthopaedic Surgeons When Diagnosing and Treating Pediatric and Adolescent Distal Radius Fractures.
Distal radius fractures are the most common injury in the pediatric population. The purpose of this study was to determine the variation among pediatric orthopaedic surgeons when diagnosing and treating distal radius fractures. Nine pediatric orthopaedic surgeons reviewed 100 sets of wrist radiographs and were asked to describe the fracture, prescribe the type of treatment and length of immobilization, and determine the next follow-up visit. κ statistics were performed to assess the agreement with the chance agreement removed. Only fair agreement was present when diagnosing and classifying the distal radius fractures (κ=0.379). There was poor agreement regarding the type of treatment that would be recommended (κ=0.059). There was no agreement regarding the length of immobilization (κ=-0.004).Poor agreement was also present regarding when the first follow-up visit should occur (κ=0.088), whether or not new radiographs should be obtained at the first follow-up visit (κ=0.133), and if radiographs were necessary at the final follow-up visit (κ=0.163). Surgeons had fair agreement regarding stability of the fracture (κ=0.320).A subgroup analysis comparing various traits of the treatment immobilization showed providers only had a slight level of agreement on whether splint or cast immobilization should be used (κ=0.072). There was poor agreement regarding whether long-arm or short-arm immobilization should be prescribed (κ=-0.067).Twenty-three of the 100 radiographs were diagnosed as a torus/buckle fracture by all 9 surgeons. κ analysis performed on all the treatment and management questions showed that each query had poor agreement. The interobserver reliability of diagnosing pediatric distal radius fractures showed only fair agreement. This study demonstrates that there is no standardization regarding how to treat these fractures and the length of immobilization required for proper fracture healing. Better classification systems of distal radius fractures are needed that standardize the treatment of these injuries. Level II.


Resistance gene cloning from a wild crop relative by sequence capture and association genetics.
Disease resistance (R) genes from wild relatives could be used to engineer broad-spectrum resistance in domesticated crops. We combined association genetics with R gene enrichment sequencing (AgRenSeq) to exploit pan-genome variation in wild diploid wheat and rapidly clone four stem rust resistance genes. AgRenSeq enables R gene cloning in any crop that has a diverse germplasm panel.


Groundwater, soil and compost, as possible sources of virulent and antibiotic-resistant Pseudomonas aeruginosa.
Pseudomonas aeruginosa is a major public health concern all around the world. In the frame of this work, a set of diverse environmental P. aeruginosa isolates with various antibiotic resistance profiles were examined in a Galleria mellonella virulence model. Motility, serotypes, virulence factors and biofilm-forming ability were also examined. Molecular types were determined by pulsed-field gel electrophoresis (PFGE). Based on our results, the majority of environmental isolates were virulent in the G. mellonella test and twitching showed a positive correlation with mortality. Resistance against several antibiotic agents such as Imipenem correlated with a lower virulence in the applied G. mellonella model. PFGE revealed that five examined environmental isolates were closely related to clinically detected pulsed-field types. Our study demonstrated that industrial wastewater effluents, composts, and hydrocarbon-contaminated sites should be considered as hot spots of high-risk clones of P. aeruginosa.


[Prevalence of dental fluorosis in children between 6 to 15 years old].
dental fluorosis is a defect in the formation of the enamel by high fluoride concentrations during tooth development. It produces hypomineralization of the enamel by increasing the porosity, thus exposing the tooth to decay. to determine the prevalence and severity of fluorosis in children aged 6 to 15 years. a sample of 456 children was evaluated. The diagnosis of fluorosis was made through physical examination and the severity of the damage was described by using a modified version of Dean's scale. 270 children presented fluorosis (59.2 %). The most affected were children of 8 to 12 years, students of first to fifth year of primary school. The severity was mild and very mild in 90 % of cases. Tooth decay appeared in 55 % of children with fluorosis and in 43 % of children without fluorosis. the prevalence of dental fluorosis is rapidly increasing. Tooth decay affected more often children with fluorosis. Assuming that the prevalence of fluorosis is due to excessive fluoride application rather than to environmental exposure is possible.


The Sivaganga eye survey: I. Blindness and cataract surgery.
To assess the prevalence of visual acuity impairment, blindness, and cataract surgery among older adults in rural southern India. Random selection of village- and urban-based clusters was used to identify a cross-sectional sample of persons 50 years of age or older from the Sivaganga district of Tamil Nadu. Subjects in 25 selected clusters were enumerated through a door-to-door survey and invited to examination sites for measurement of uncorrected, presenting, and best-corrected visual acuity and ocular examination in 1999. The principal cause was identified for eyes with presenting visual acuity worse than 6/18. Quality assurance monitoring of visual acuity measurements took place in five of the study clusters. A total of 5081 persons in 3517 households were enumerated, and 4642 (91.4%) were examined. Thirty-six percent presented with visual acuity worse than 6/18 in the better eye. The prevalence of blindness, based on visual acuity worse than 6/60 in both eyes, was 6.0% (95% confidence interval [CI]: 5.1% to 6.9%) with presenting vision, and 2.5% (95% CI: 1.8% to 3.1%) with best correction. Blindness with presenting visual acuity was associated with older age and illiteracy. Cataract was the principal cause of blindness in one or both eyes in 69.4% of those presenting blind, and uncorrected aphakia and other refractive error affected 35.6% in at least one eye. The prevalence of cataract surgery was 14.7% (95% CI: 13.0% to 16.4%); low surgical coverage among the cataract blind was associated with illiteracy. It appears that much has been done in the prevention of blindness in Sivaganga. Nevertheless, blindness remains an important public health problem, mainly because of cataract and refractive error. Prevention of blindness programs in the area should target these two causes, with special emphasis on the elderly and the illiterate.


Atherogenic dyslipidemia and oxidative stress: a new look.
Although results from in vitro studies and clinical trials demonstrate strong associations between oxidative stress and cardiovascular risk, to date still no convincing data are available to suggest that treatment with antioxidants might reduce vascular events. Oxidative modifications of low-density lipoproteins (LDL) represent an early stage of atherosclerosis, and small, dense LDL are more susceptible to oxidation than larger, more buoyant particles. Oxidized LDL are independent predictors of subclinical and clinical atherosclerosis. Recent studies suggested that novel therapeutic strategies may take into account the removal of such particles from circulation. Future research is required to explore the potential synergistic impact of markers of oxidative stress and atherogenic dyslipidemia, particularly small dense LDL, on cardiovascular risk.


Molecular identification and expressive characterization of an olfactory co-receptor gene in the Asian honeybee, Apis cerana cerana.
Olfaction recognition process is extraordinarily complex in insects, and the olfactory receptors play an important function in the process. In this paper, a highly conserved olfactory co-receptor gene, AcerOr2 (ortholog to the Drosophila melanogaster Or83b), cloned from the antennae of the Asian honeybee, Apis cerana cerana Fabricius (Hymenoptera: Apidae), using reverse transcriptase PCR and rapid amplification of cDNA ends. The full-length sequence of the gene was 1763 bp long, and the cDNA open reading frame encoded 478 amino acid residues, including 7 putative transmembrane domains. Alignment analysis revealed that AcerOr2 shares high homology (> 74%) with similar olfactory receptors found in other Hymenoptera species. The amino acid identity with the closely related species Apis mellifera reached 99.8%. The developmental expression analysis using quantitative real-time reverse transcriptase PCR suggested that the AcerOr2 transcript was expressed at a relatively low level in the larval stage, whereas it was expressed broadly in the pupal and adult stages, with a significantly high level on the days just before and after eclosion. In situ hybridization showed that AcerOr2 mRNA was expressed in sensilla placodea and on the basal region of the worker antennal cuticle, in accordance with the previous conclusions that the conserved genes are expressed in most olfactory receptor neurons.


Trust makers, breakers and brokers: building trust in the Australian food system.
The importance of consumer trust in the food supply has previously been identified, and dimensions of consumer trust in food-who they trust and the type of trust that they exhibit-has been explored. However, there is a lack of research about the mechanisms through which consumer trust in the food supply is developed, maintained, broken and repaired. This study seeks to address this gap by exploring if, and how, consumer trust in the food supply is considered by the media, food industry and governments when responding to food scares. The aim of the research is to develop models of trust building that can be implemented following food scares. Semi-structured interviews will be undertaken with media, public relations officials and policy makers in Australia, New Zealand and the United Kingdom. Participants will be recruited through purposive sampling and will be asked to discuss a hypothetical case study outlining a food incident, and any experiences of specific food scares. Models of trust development, maintenance and repair will be developed from interview data. Comment on these models will be sought from experts in food-related organizations through a Delphi study, where participants will be asked to consider the usefulness of the models. Participants' comments will be used to revise the models until consensus is reached on the suitability and usability of the models. This study will contribute to the literature about systems-based trust, and explore trust as a social and regulatory process. The protocol and results will be of interest and use to the food industry, food regulators, consumer advocate groups, media seeking to report food-related issues and policy makers concerned with public health and consumer health and well-being. This research represents an important contribution to the translation of the theoretical conceptualizations of trust into practical use in the context of food.


The Role of Bystander Effects in the Antitumor Activity of the Hypoxia-Activated Prodrug PR-104.
Activation of prodrugs in tumors (e.g., by bioreduction in hypoxic zones) has the potential to generate active metabolites that can diffuse within the tumor microenvironment. Such "bystander effects" may offset spatial heterogeneity in prodrug activation but the relative importance of this effect is not understood. Here, we quantify the contribution of bystander effects to antitumor activity for the first time, by developing a spatially resolved pharmacokinetic/pharmacodynamic (SR-PK/PD) model for PR-104, a phosphate ester pre-prodrug that is converted systemically to the hypoxia-activated prodrug PR-104A. Using Green's function methods we calculated concentrations of oxygen, PR-104A and its active metabolites, and resultant cell killing, at each point of a mapped three-dimensional tumor microregion. Model parameters were determined in vitro, using single cell suspensions to determine relationships between PR-104A metabolism and clonogenic cell killing, and multicellular layer (MCL) cultures to measure tissue diffusion coefficients. LC-MS/MS detection of active metabolites in the extracellular medium following exposure of anoxic single cell suspensions and MCLs to PR-104A confirmed that metabolites can diffuse out of cells and through a tissue-like environment. The SR-PK/PD model estimated that bystander effects contribute 30 and 50% of PR-104 activity in SiHa and HCT116 tumors, respectively. Testing the model by modulating PR-104A-activating reductases and hypoxia in tumor xenografts showed overall clonogenic killing broadly consistent with model predictions. Overall, our data suggest that bystander effects are important in PR-104 antitumor activity, although their reach may be limited by macroregional heterogeneity in hypoxia and reductase expression in tumors. The reported computational and experimental techniques are broadly applicable to all targeted anticancer prodrugs and could be used to identify strategies for rational prodrug optimization.


Effects of clinical and environmental factors on bronchoalveolar antibody responses to Pneumocystis jirovecii: A prospective cohort study of HIV+ patients.
Humoral immunity plays an important role against Pneumocystis jirovecii infection, yet clinical and environmental factors that impact bronchoalveolar antibody responses to P. jirovecii remain uncertain. From October 2008-December 2011 we enrolled consecutive HIV-infected adults admitted to San Francisco General Hospital (SFGH) who underwent bronchoscopy for suspected Pneumocystis pneumonia (PCP). We used local air quality monitoring data to assign ozone, nitrogen dioxide, and fine particulate matter exposures within 14 days prior to hospital admission. We quantified serum and bronchoalveolar lavage fluid (BALF) antibody responses to P. jirovecii major surface glycoprotein (Msg) recombinant constructs using ELISA. We then fit linear regression models to determine whether PCP and ambient air pollutants were associated with bronchoalveolar antibody responses to Msg. Of 81 HIV-infected patients enrolled, 47 (58%) were diagnosed with current PCP and 9 (11%) had a prior history of PCP. The median CD4+ count was 51 cells/μl (IQR 15-129) and 44% were current smokers. Serum antibody responses to Msg were statistically significantly predictive of BALF antibody responses, with the exception of IgG responses to MsgC8 and MsgC9. Prior PCP was associated with increased BALF IgA responses to Msg and current PCP was associated with decreased IgA responses. For instance, among patients without current PCP, those with prior PCP had a median 73.2 U (IQR 19.2-169) IgA response to MsgC1 compared to a 5.00 U (3.52-12.6) response among those without prior PCP. Additionally, current PCP predicted a 22.5 U (95%CI -39.2, -5.82) lower IgA response to MsgC1. Ambient ozone within the two weeks prior to hospital admission was associated with decreased BALF IgA responses to Msg while nitrogen dioxide was associated with increased IgA responses. PCP and ambient air pollutants were associated with BALF IgA responses to P. jirovecii in HIV-infected patients evaluated for suspected PCP.


Enhanced dopaminergic neurotoxicity mediated by MPTP in IL-32β transgenic mice.
Parkinson's disease (PD) is a neurodegenerative disorder characterized by prominent loss of the nigral dopaminergic neurons and motor symptoms, such as resting tremor and bradykinesia. Evidence suggests that neuroinflammation may play a critical role in PD pathogenesis. Interleukin (IL)-32 is a newly-identified proinflammatory cytokine, which regulates innate and adaptive immune responses by activating p38 MAPK and NF-κB signaling pathways. The cytokine has been implicated in cancers and autoimmune, inflammatory, and infectious diseases. In this study, we attempted to identify the effects of IL-32β on dopaminergic neurotoxicity induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), using IL-32β transgenic mice. Male wild type and IL-32β transgenic mice received intraperitoneal injections of vehicle or MPTP (15 mg/kg × 4). Immunohistochemistry showed that overexpression of IL-32β significantly increased MPTP-mediated loss of dopaminergic neurons in the substantia nigra and deletion of tyrosine hydroxylase-positive fibers in the striatum. Dopamine depletion in the striatum and deficit in locomotor activity were enhanced in IL-32β transgenic mice. These results were accompanied by higher neuroinflammatory responses in the brains of transgenic mice. Finally, we found that IL-32β exaggerated MPTP-mediated activation of p38 MAPK and JNK pathways, which have been shown to be involved in MPTP neurotoxicity. These results suggest that IL-32β exacerbates MPTP neurotoxicity through enhanced neuroinflammatory responses.


Pre-PCI versus immediate post-PCI Impella initiation in acute myocardial infarction complicated by cardiogenic shock.
In selected patients with an acute myocardial infarction (AMI) complicated by Cardiogenic shock (CS), mechanical circulatory support with Impella may be beneficial, although conclusive evidence is still lacking. Nevertheless, it has been suggested that Impella initiation prior to primary PCI might improve survival. To investigate the effect pre-PCI versus immediate post-PCI Impella initiation on short term mortality. A prospective, single center, observational study, was performed including all patients with STEMI complicated by CS, treated with primary PCI and Impella. Thirty day mortality was compared between patients with Impella initiation pre-PCI and immediately post-PCI. A total of 88 patients were included. In the pre-PCI group (n = 21), admission heart rate was lower (84 versus 94 bpm, p = 0.04) and no IABP was implanted before Impella initiation, versus 17.9% in post-PCI group (n = 67), p = 0.04. Total 30-day all-cause mortality was 58%, and was lower in pre-PCI group, 47.6% versus 61.2% in the post-PCI group, however not statistically significant (HR 0.7, 95% CI 0.3-1.3, p = 0.21). Thirty-day cardiac mortality was significantly lower in the pre-PCI group, 19% versus 44.7% in the post-PCI group (HR 0.3, 95% CI 0.09-0.96, p = 0.042). Pre-PCI Impella initiation in AMICS patients was not associated with a statistically significant difference in 30-day all-cause mortality, compared to post-PCI Impella initiation.


Ecosystem ecology meets adaptive management: food web response to a controlled flood on the Colorado River, Glen Canyon.
Large dams have been constructed on rivers to meet human demands for water, electricity, navigation, and recreation. As a consequence, flow and temperature regimes have been altered, strongly affecting river food webs and ecosystem processes. Experimental high-flow dam releases, i.e., controlled floods, have been implemented on the Colorado River, U.S.A., in an effort to reestablish pulsed flood events, redistribute sediments, improve conditions for native fishes, and increase understanding of how dam operations affect physical and biological processes. We quantified secondary production and organic matter flows in the food web below Glen Canyon dam for two years prior and one year after an experimental controlled flood in March 2008. Invertebrate biomass and secondary production declined significantly following the flood (total biomass, 55% decline; total production, 56% decline), with most of the decline driven by reductions in two nonnative invertebrate taxa, Potamopyrgus antipodarum and Gammarus lacustris. Diatoms dominated the trophic basis of invertebrate production before and after the controlled flood, and the largest organic matter flows were from diatoms to the three most productive invertebrate taxa (P. antipodarum, G. lacustris, and Tubificida). In contrast to invertebrates, production of rainbow trout (Oncorhynchus mykiss) increased substantially (194%) following the flood, despite the large decline in total secondary production of the invertebrate assemblage. This counterintuitive result is reconciled by a post-flood increase in production and drift concentrations of select invertebrate prey (i.e., Chironomidae and Simuliidae) that supported a large proportion of trout production but had relatively low secondary production. In addition, interaction strengths, measured as species impact values, were strongest between rainbow trout and these two taxa before and after the flood, demonstrating that the dominant consumer-resource interactions were not necessarily congruent with the dominant organic matter flows. Our study illustrates the value of detailed food web analysis for elucidating pathways by which dam management may alter production and strengths of species interactions in river food webs. We suggest that controlled floods may increase production of nonnative rainbow trout, and this information can be used to help guide future dam management decisions.


Transcription initiation patterns indicate divergent strategies for gene regulation at the chromatin level.
The application of deep sequencing to map 5' capped transcripts has confirmed the existence of at least two distinct promoter classes in metazoans: "focused" promoters with transcription start sites (TSSs) that occur in a narrowly defined genomic span and "dispersed" promoters with TSSs that are spread over a larger window. Previous studies have explored the presence of genomic features, such as CpG islands and sequence motifs, in these promoter classes, but virtually no studies have directly investigated the relationship with chromatin features. Here, we show that promoter classes are significantly differentiated by nucleosome organization and chromatin structure. Dispersed promoters display higher associations with well-positioned nucleosomes downstream of the TSS and a more clearly defined nucleosome free region upstream, while focused promoters have a less organized nucleosome structure, yet higher presence of RNA polymerase II. These differences extend to histone variants (H2A.Z) and marks (H3K4 methylation), as well as insulator binding (such as CTCF), independent of the expression levels of affected genes. Notably, differences are conserved across mammals and flies, and they provide for a clearer separation of promoter architectures than the presence and absence of CpG islands or the occurrence of stalled RNA polymerase. Computational models support the stronger contribution of chromatin features to the definition of dispersed promoters compared to focused start sites. Our results show that promoter classes defined from 5' capped transcripts not only reflect differences in the initiation process at the core promoter but also are indicative of divergent transcriptional programs established within gene-proximal nucleosome organization.


Making "real" molecules in virtual space.
Predicting "realistic" compounds of given chemical reactions with virtual synthesis tools usually requires the manual intervention of experienced chemists in the enumeration phase for the selection of appropriate reactants, assignment of the corresponding reaction sites, and removal of the unlikely products. To automate the virtual synthesis process, we have moved the expertise intensive parts from the compound library design phase to the reaction library design phase. ChemAxon is building an in silico reaction library containing important preparative transformations, where each reaction definition contains a generic transformation scheme and additional rules to handle the various starting compounds according to the corresponding chemo-, regio-, and stereoselectivity issues. Having well designed reaction definitions in hand, our software tool is able to generate synthetically feasible compound libraries with minimal effort in the enumeration phase.


Immunogenic salivary proteins of Triatoma infestans: development of a recombinant antigen for the detection of low-level infestation of triatomines.
Triatomines are vectors of Trypanosoma cruzi, the etiological agent of Chagas disease in Latin America. The most effective vector, Triatoma infestans, has been controlled successfully in much of Latin America using insecticide spraying. Though rarely undertaken, surveillance programs are necessary in order to identify new infestations and estimate the intensity of triatomine bug infestations in domestic and peridomestic habitats. Since hosts exposed to triatomines develop immune responses to salivary antigens, these responses can be evaluated for their usefulness as epidemiological markers to detect infestations of T. infestans. T. infestans salivary proteins were separated by 2D-gel electrophoresis and tested for their immunogenicity by Western blotting using sera from chickens and guinea pigs experimentally exposed to T. infestans. From five highly immunogenic protein spots, eight salivary proteins were identified by nano liquid chromatography-electrospray ionization-tandem mass spectrometry (nanoLC-ESI-MS/MS) and comparison to the protein sequences of the National Center for Biotechnology Information (NCBI) database and expressed sequence tags of a unidirectionally cloned salivary gland cDNA library from T. infestans combined with the NCBI yeast protein sub-database. The 14.6 kDa salivary protein [gi|149689094] was produced as recombinant protein (rTiSP14.6) in a mammalian cell expression system and recognized by all animal sera. The specificity of rTiSP14.6 was confirmed by the lack of reactivity to anti-mosquito and anti-sand fly saliva antibodies. However, rTiSP14.6 was recognized by sera from chickens exposed to four other triatomine species, Triatoma brasiliensis, T. sordida, Rhodnius prolixus, and Panstrongylus megistus and by sera of chickens from an endemic area of T. infestans and Chagas disease in Bolivia. The recombinant rTiSP14.6 is a suitable and promising epidemiological marker for detecting the presence of small numbers of different species of triatomines and could be developed for use as a new tool in surveillance programs, especially to corroborate vector elimination in Chagas disease vector control campaigns.


Supershrimp: deep bioturbation in the strait of canso, nova scotia.
Axius serratus, a crustacean thought to be extremely rare, was discovered in large numbers in polluted regions of the Strait of Canso. The shrimp may live deeper than 3 meters in the sediment; burrows are kept open to at least 2.5 meters. Sediment contained in old filled burrows is anomalous in its distribution of particle size and its content of water, organic carbon, and trace elements. These anomalous qualitites affect the geotechnical properties of sediments on the sea floor.


Contrasuppression in autoimmunity. Abnormal contrasuppression facilitates expression of nephritogenic effector T cells and interstitial nephritis in kdkd mice.
We have used the murine model of spontaneous autoimmune interstitial nephritis in kdkd mice to examine the importance of abnormal immunoregulation in the expression of disease. T cells from naive congenic CBA/Ca mice suppress both histologic renal injury in the kdkd strain as well as the DTH reactivity to CBA/Ca renal tubular antigens mediated by lymphocytes from nephritic kdkd mice. These antigen-specific suppressor T cells are Lyt-2+, L3T4+, I-Jk+, genetically dominant and I-Jk restricted. Unfractionated spleen cells from young, prenephritic kdkd mice also demonstrate such suppressor function. Shortly preceding disease onset, however, net suppression is functionally bypassed by emergent contrasuppressor T cells. These regulatory cells are also Lyt-2+ and I-Jk+, and adhere both to the Vicia Villosa lectin and CBA/Ca TBM. By admixing these contrasuppressor cells with spleen cells from non-disease-prone CBA/Ca mice we were able to demonstrate the presence of DTH-reactive and nephritogenic effector cells in the latter population. Such nephritogenic effector cells could also be simply demonstrated after depletion of the suppressor cells with anti-I-Jk mAbs and complement. These findings support a role for contrasuppressor cells in the abrogation of tolerance to parenchymal self-antigens.


Inhibitors of sterol synthesis. Synthesis and spectral properties of 3 beta-hydroxy-25,26,26,26,27,27,27-heptafluoro-5 alpha-cholestan-15-one.
3 beta-Hydroxy-25,26,26,26,27,27,27-heptafluoro-5 alpha-cholestan-15-one (4) has been prepared as part of a program to synthesize 15-ketosterols that are not readily metabolized to cholesterol or side-chain oxygenated species. Saponification of 3 beta-acetoxy-5 alpha-chola-8(14),23-dien-15-one (5) followed by lithium-ammonia reduction with a bromobenzene quench gave 3 beta-hydroxy-5 alpha-chol-23-en-15-one (6). Addition of (CF3)2CFI to 6 in the presence of triethylborane gave an iodide preparation, which was reduced to 4 with tributyltin hydride (71% overall yield of 4 from 5). The 23-iodide preparations consisted of 6:1 mixtures of (23R)-3 beta-hydroxy-23-iodo-25,26,26,26,27,27,27-heptafluoro-5 alpha-cholestan-15-one (9a) and its C-23 epimer 9b with variable amounts of 4. Compound 4 was also prepared by lithium-ammonia reduction of the delta 8(14) analogs of 4 and iodides 9a and 9b. The presence of small amounts of 6 in the latter product suggested a side reaction involving cleavage of the C24-C25 bond with loss of a (CF3)2CF radical. Also prepared were 25,26,26,26,27,27,27-heptafluoro-5 alpha-cholestane-3 beta, 15 alpha-diol, its 15 beta epimer, the 7 alpha-methyl analog of 4, 3 beta-hydroxy-7 alpha-methyl-5 alpha-cholestan-15-one (16), and (25R)-3 beta,26-dihydroxy-5 alpha-cholestan-15-one. Full 1H and 13C-NMR data of high precision with complete signal assignments are given for all new compounds. Definitive 1H-NMR stereochemical assignments of the C-24 protons were established for most sterols with a C8H17 side chain based on analysis of the downfield H-24 resonance in a 750-MHz spectrum of 16. Detailed electron-impact mass spectral data are presented together with a summary of major fragmentation patterns for 15-hydroxy- and 15-ketosteroids with and without a delta 8(14) bond.


Data Generated by Quantitative Liquid Chromatography-Mass Spectrometry Proteomics Are Only the Start and Not the Endpoint: Optimization of Quantitative Concatemer-Based Measurement of Hepatic Uridine-5'-Diphosphate-Glucuronosyltransferase Enzymes with Reference to Catalytic Activity.
Quantitative proteomic methods require optimization at several stages, including sample preparation, liquid chromatography-tandem mass spectrometry (LC-MS/MS), and data analysis, with the final analysis stage being less widely appreciated by end-users. Previously reported measurement of eight uridine-5'-diphospho-glucuronosyltransferases (UGT) generated by two laboratories [using stable isotope-labeled (SIL) peptides or quantitative concatemer (QconCAT)] reflected significant disparity between proteomic methods. Initial analysis of QconCAT data showed lack of correlation with catalytic activity for several UGTs (1A4, 1A6, 1A9, 2B15) and moderate correlations for UGTs 1A1, 1A3, and 2B7 (Rs = 0.40-0.79, P < 0.05; R2 = 0.30); good correlations were demonstrated between cytochrome P450 activities and abundances measured in the same experiments. Consequently, a systematic review of data analysis, starting from unprocessed LC-MS/MS data, was undertaken, with the aim of improving accuracy, defined by correlation against activity. Three main criteria were found to be important: choice of monitored peptides and fragments, correction for isotope-label incorporation, and abundance normalization using fractional protein mass. Upon optimization, abundance-activity correlations improved significantly for six UGTs (Rs = 0.53-0.87, P < 0.01; R2 = 0.48-0.73); UGT1A9 showed moderate correlation (Rs = 0.47, P = 0.02; R2 = 0.34). No spurious abundance-activity relationships were identified. However, methods remained suboptimal for UGT1A3 and UGT1A9; here hydrophobicity of standard peptides is believed to be limiting. This commentary provides a detailed data analysis strategy and indicates, using examples, the significance of systematic data processing following acquisition. The proposed strategy offers significant improvement on existing guidelines applicable to clinically relevant proteins quantified using QconCAT.


Hemostatic markers in surgery: a different fibrinolytic activity may be of pathophysiological significance in orthopedic versus abdominal surgery.
Without prophylaxis, patients subjected to major abdominal surgery have a risk of deep vein thrombosis of approximately 30%, while the rate varies between 40% and 60% in orthopedic surgery. The reasons for this discrepancy are not completely understood. The present study was designed to compare the pre- and postoperative behavior of different coagulation and fibrinolysis parameters in patients undergoing both types of surgery, receiving low molecular weight heparin prophylaxis. Samples were taken before operation and on postoperative days 1, 3, and 7. The following parameters were assessed: prothrombin fragment 1 + 2, thrombin-antithrombin III complexes, fibrinopeptide A, tissue plasminogen activator, plasminogen activator inhibitor, plasmin-alpha 2-antiplasmin complexes, and fibrin degradation products. We found a significant increase in the clotting markers postoperatively compared with preoperative values (P < 0.05), both in abdominal and orthopedic surgery, indicating a marked hemostatic activation which remained until postoperative day 7. A significant increase in plasminogen activator inhibitor (P < 0.01) and a decrease in tissue plasminogen activator and plasmin-alpha 2-antiplasmin complexes was also observed early after operation. The plasminogen activator inhibitor activity decreased, while tissue plasminogen activator and plasmin-alpha 2-antiplasmin levels increased significantly on days 3 and 7 (P < 0.05). Fibrin degradation products significantly increased throughout the postoperative period (P < 0.01). Preoperatively, we found higher plasminogen activator inhibitor activity and lower tissue plasminogen activator and plasmin-alpha 2-antiplasmin complexes (P < 0.05) in patients undergoing hip replacement compared with abdominal surgery. Fibrin degradation products were also significantly lower on postoperative day 3 in patients undergoing hip replacement (P < 0.01). We suggest that the lower preoperative fibrinolytic activation observed in patients undergoing orthopedic surgery compared with abdominal surgery might have pathophysiological consequences. Our results also indicate that the hemostatic activation persists beyond the 7th postoperative day despite prophylaxis.


Cytokine Storm in COVID-19-Immunopathological Mechanisms, Clinical Considerations, and Therapeutic Approaches: The REPROGRAM Consortium Position Paper.
Cytokine storm is an acute hyperinflammatory response that may be responsible for critical illness in many conditions including viral infections, cancer, sepsis, and multi-organ failure. The phenomenon has been implicated in critically ill patients infected with SARS-CoV-2, the novel coronavirus implicated in COVID-19. Critically ill COVID-19 patients experiencing cytokine storm are believed to have a worse prognosis and increased fatality rate. In SARS-CoV-2 infected patients, cytokine storm appears important to the pathogenesis of several severe manifestations of COVID-19: acute respiratory distress syndrome, thromboembolic diseases such as acute ischemic strokes caused by large vessel occlusion and myocardial infarction, encephalitis, acute kidney injury, and vasculitis (Kawasaki-like syndrome in children and renal vasculitis in adult). Understanding the pathogenesis of cytokine storm will help unravel not only risk factors for the condition but also therapeutic strategies to modulate the immune response and deliver improved outcomes in COVID-19 patients at high risk for severe disease. In this article, we present an overview of the cytokine storm and its implications in COVID-19 settings and identify potential pathways or biomarkers that could be targeted for therapy. Leveraging expert opinion, emerging evidence, and a case-based approach, this position paper provides critical insights on cytokine storm from both a prognostic and therapeutic standpoint.


Racial Non-equivalence of Socioeconomic Status and Self-rated Health among African Americans and Whites.
Racial health inequities are not fully explained by socioeconomic status (SES) measures like education, income and wealth. The largest inequities are observed among African American and white college graduates suggesting that African Americans do not receive the same health benefits of education. African Americans do not receive the same income and wealth returns of college education as their white counterparts indicating a racial non-equivalence of SES that may affect health inequities. The aim of this study is to determine whether racial non-equivalence of SES mediates race inequities in self-rated health by education and sex. Using data from the 2007-2016 National Health and Nutrition Examination Survey in the United States, the mediation of the associations between race and self-rated health through household income ≥400% federal poverty line, homeownership, and investment income were assessed among college graduates and non-college graduates by sex. Indirect associations were observed among college graduate women (odds = 0.08, standard error (s.e.) = 0.03), and non-college graduate men (odds = 0.14, s.e. = 0.02) and women (odds = 0.06, s.e. = 0.02). Direct associations between race and self-rated health remained after accounting for household income and wealth indicators suggesting that race differences in income and wealth partially mediate racial inequities in self-rated health. This study demonstrates that the racial non-equivalence of SES has implications for health inequities, but the magnitude of indirect associations varied by sex. Other factors like discrimination, health pessimism and segregation should be considered in light of the racial non-equivalence of SES and racial inequities in self-rated health.


Ranbp2 haploinsufficiency mediates distinct cellular and biochemical phenotypes in brain and retinal dopaminergic and glia cells elicited by the Parkinsonian neurotoxin, 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP).
Many components and pathways transducing multifaceted and deleterious effects of stress stimuli remain ill-defined. The Ran-binding protein 2 (RanBP2) interactome modulates the expression of a range of clinical and cell-context-dependent manifestations upon a variety of stressors. We examined the role of Ranbp2 haploinsufficiency on cellular and metabolic manifestations linked to tyrosine-hydroxylase (TH(+)) dopaminergic neurons and glial cells of the brain and retina upon acute challenge to 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), a parkinsonian neurotoxin, which models facets of Parkinson disease. MPTP led to stronger akinetic parkinsonism and slower recovery in Ranbp2 (+/-) than wild-type mice without viability changes of brain TH(+)-neurons of either genotype, with the exception of transient nuclear atypia via changes in chromatin condensation of Ranbp2 (+/-) TH(+)-neurons. Conversely, the number of wild-type retinal TH(+)-amacrine neurons compared to Ranbp2 (+/-) underwent milder declines without apoptosis followed by stronger recoveries without neurogenesis. These phenotypes were accompanied by a stronger rise of EdU(+)-proliferative cells and non-proliferative gliosis of GFAP(+)-Müller cells in wild-type than Ranbp2 (+/-) that outlasted the MPTP-insult. Finally, MPTP-treated wild-type and Ranbp2 (+/-) mice present distinct metabolic footprints in the brain or selective regions thereof, such as striatum, that are supportive of RanBP2-mediated regulation of interdependent metabolic pathways of lysine, cholesterol, free-fatty acids, or their β-oxidation. These studies demonstrate contrasting gene-environment phenodeviances and roles of Ranbp2 between dopaminergic and glial cells of the brain and retina upon oxidative stress-elicited signaling and factors triggering a continuum of metabolic and cellular manifestations and proxies linked to oxidative stress, and chorioretinal and neurological disorders such as Parkinson.


Molecular Pathways Associated With Methylmercury-Induced Nrf2 Modulation.
Methylmercury (MeHg) is a potent neurotoxin that affects particularly the developing brain. Since MeHg is a potent electrophilic agent, a wide range of intracellular effects occur in response to its exposure. Yet, the molecular mechanisms associated with MeHg-induced cell toxicity have yet to be fully understood. Activation of cell defense mechanisms in response to metal exposure, including the up-regulation of Nrf2- (nuclear factor erythroid 2-related factor 2)-related genes has been previously shown. Nrf2 is a key regulator of cellular defenses against oxidative, electrophilic and environmental stress, regulating the expression of antioxidant proteins, phase-II xenobiotic detoxifying enzymes as well phase-III xenobiotic transporters. Analogous to other electrophiles, MeHg activates Nrf2 through modification of its repressor Keap1 (Kelch-like ECH-associated protein 1). However, recent findings have also revealed that Keap1-independent signal pathways might contribute to MeHg-induced Nrf2 activation and cytoprotective responses against MeHg exposure. These include, Akt phosphorylation (Akt/GSK-3β/Fyn-mediated Nrf2 activation pathway), activation of the PTEN/Akt/CREB pathway and MAPK-induced autophagy and p62 expression. In this review, we summarize the state-of-the-art knowledge regarding Nrf2 up-regulation in response to MeHg exposure, highlighting the modulation of signaling pathways related to Nrf2 activation. The study of these mechanisms is important in evaluating MeHg toxicity in humans, and can contribute to the identification of the molecular mechanisms associated with MeHg exposure.


Cancer profile of Hyderabad, Pakistan 1998-2002.
Hyderabad is the third largest city of Pakistan, the second largest city of Sindh Province and one of the oldest cities of the sub-continent. This administrative headquarter is located just east of the River Indus and is an important commercial and industrial center. Once a provincial capital, it is at a distance of approximately 200-km from Karachi. This present study was conducted with the objective of providing the cancer profile of Hyderabad, which has an urban population of 2,840,653 (52.2% M, 47.8% F) annual growth rate 1.13. The city is inhabited by all ethnicities of the country, however the predominant ethnicity is Sindhi, followed by Mohajirs (post-partition immigrants from India), and a lesser extent other ethnicities of Pakistan viz. Baluchs, Punjabis and Pathans. The study includes two sets of patients. First the incident cancer cases, residents of Hyderabad, who reached Karachi for diagnosis or treatment. Second the incident cancer cases registered at the Aga Khan University Pathology-based Cancer Registry (APCR) Pathology collection points at Hyderabad and subsequently registered at APCR, during 1st January 1998 to 31st December 2002. The pathology department of the AKU has 3 centers in Hyderabad, which provide diagnostic pathology especially oncopathology services to the city. The age-standardized rates (ASR) for cancer (all sites) 1998 to 2002 in Hyderabad were 91.6/100,000 in males and 96.0/100,000 in females. The most common malignancies (ASR per 100,000) in males were oral cavity (11.8), lymphoma (10.6), lung (8.0), urinary bladder (6.8), prostate (4.8), liver (4.4), pharynx (4.2), colo-rectum (3.6), larynx (3.2), and skin (3.2). The cancers in females (ASR per 100,000) were breast (22.4), oral cavity (11.5), gall bladder (4.8), esophagus (4.2), cervix (3.6), ovary (3.4), colo-rectum (3.4), lymphoma (3.4), uterus (3.4), and thyroid (2.4). Tobacco-associated cancers were responsible for approximately 40.0% of the tumors in males and 20.0% in females. Histological confirmation remained 96.3%, with 44.5% presenting in grade II or I, 55.5% presenting as stage III and IV. Information on grade and stage of malignancy was available in 70% and 50% of the cases respectively. Males comprised 53.1%, and females 46.9% of the cases. The mean age of cancer all sites, both genders was 45.2 years (95% CI 44.4; 45.9), males 45.4 years (95% CI 44.3; 46.5); females 44.9 years (95% CI 43.9; 45.9). Conclusions drawn from this database must be interpreted with care, as it may be identified as data from selected medical institutions. Chances of selective collection bias are minimized as the data of the AKU pathology is collected from multiple centers in the city of Hyderabad, dispersed at distances, which allows adequate sampling from the entire city. There is a slight preponderance of lymphomas in males which we feel is a true higher risk, yet it may indicate an over representation of easily accessible sites in pathology based-data. Nonetheless, this is the first attempt to determine the cancer incidence pattern of Hyderabad, and should serve as a guideline for estimation of the cancer burden and risk assessment statistics of Pakistan and the cancer control program of the country.


Prevalence of depression and associated risk factors among the elderly in urban and rural field practice areas of a tertiary care institution in Ludhiana.
Depression, the most common psychiatric disorder among the elderly, is not yet perceived as an important health problem in India, where few population-based studies have addressed this problem. To estimate the prevalence of depression and identify the associated risk factors in the elderly population. 3038 consenting elderly (>60 years old) rural and urban residents of both sexes from the field practice areas were interviewed and examined in a cross-sectional study. Physical impairment in the subjects was assessed with the Everyday Abilities Scale for India (EASI), depression by the 15-item Geriatric Depression Scale (GDS-15), and cognitive impairment by the Mini-Mental State Examination (MMSE). Data were analyzed using Epi Info version-6 software. Statistical analysis included proportions, χ[2] -test, odds ratio, and its 95% confidence interval. Multiple logistic regression was done using SPSS version 21. The prevalence of depression in the study population was 8.9%. It was significantly higher in urban residents, females, older elderly, nuclear families, in those living alone, those not working, illiterates, poor, functionally impaired, and cognitively impaired. In the multivariate analysis, unmarried/widowed status, unemployment, and illiteracy did not emerge as risk factors. Urban residence, female gender, higher age, nuclear family, poverty, and functional and cognitive impairment were found to be associated with depression even after controlling for other factors.


The perceived importance of physical activity: associations with psychosocial and health-related outcomes.
The purpose of this study was to assess the extent to which participation in a 12-month exercise program changed the degree of importance that older adults attached to physical activity. In addition, associations among changes in physical activity importance and health-related and psychosocial outcomes were examined. Community-dwelling older adults (N = 179) were recruited to participate in a 12-month exercise trial examining the association between changes in physical activity and fitness with changes in brain structure and psychological health. Participants were randomly assigned to a walking condition or a flexibility, toning, and balance condition. Physical, psychological, and cognitive assessments were taken at months 0, 6, and 12. Involvement in a 12-month exercise program increased the importance that participants placed on physical activity; this positive change was similar across exercise condition and sex. Changes in importance, however, were only associated with changes in physical health status and outcome expectations for exercise midway through the intervention. There were no significant associations at the end of the program. Regular participation in physical activity can positively influence the perceived importance of the behavior itself. Yet, the implications of such changes on physical activity-related outcomes remain equivocal and warrant further investigation.


Comparison of genetic maps for two Leptospira interrogans serovars provides evidence for two chromosomes and intraspecies heterogeneity.
Genetic maps were constructed for Leptospira interrogans serovars icterohaemorrhagiae and pomona. Previously we independently constructed physical maps of the genomes for these two serovars. The genomes of both serovars consist of a large replicon (4.4 to 4.6 Mb) and a small replicon (350 kb). Genes were localized on the physical maps by using Southern blot analysis with specific probes. Among the probes used were genes encoding a variety of essential enzymes and genes usually found near bacterial chromosomal replication origins. Most of the essential genes are on the larger replicon of each serovar. However, the smaller replicons of both serovars contain the asd gene. The asd gene encodes aspartate beta-semialdehyde dehydrogenase, an enzyme essential in amino acid and cell wall biosyntheses. The finding that both L. interrogans replicons contain essential genes suggests that both replicons are chromosomes. Comparison of the genetic maps of the larger replicons of the two serovars showed evidence of large rearrangements. These data show that there is considerable intraspecies heterogeneity in L. interrogans.


Genomics in research and health care with Aboriginal and Torres Strait Islander peoples.
Genomics is increasingly becoming an integral component of health research and clinical care. The perceived difficulties associated with genetic research involving Aboriginal and Torres Strait Islander people mean that they have largely been excluded as research participants. This limits the applicability of research findings for Aboriginal and Torres Strait Islander patients. Emergent use of genomic technologies and personalised medicine therefore risk contributing to an increase in existing health disparities unless urgent action is taken. To allow the potential benefits of genomics to be more equitably distributed, and minimise potential harms, we recommend five actions: (1) ensure diversity of participants by implementing appropriate protocols at the study design stage; (2) target diseases that disproportionately affect disadvantaged groups; (3) prioritise capacity building to promote Indigenous leadership across research professions; (4) develop resources for consenting patients or participants from different cultural and linguistic backgrounds; and (5) integrate awareness of issues relating to Indigenous people into the governance structures, formal reviews, data collection protocols and analytical pipelines of health services and research projects.


Damage to temporoparietal cortex is sufficient for impaired semantic control.
Semantic control allows us to focus semantic activation on currently relevant aspects of knowledge, even in the face of competition or when the required information is weakly encoded. Diverse cortical regions, including left prefrontal and posterior temporal cortex, are implicated in semantic control, however; the relative contribution of these regions is unclear. For the first time, we compared semantic aphasia (SA) patients with damage restricted to temporoparietal cortex (TPC; N = 8) to patients with infarcts encompassing prefrontal cortex (PF+; N = 22), to determine if prefrontal lesions are necessary for semantic control deficits. These SA groups were also compared with semantic dementia (SD; N = 10), characterised by degraded semantic representations. We asked whether TPC cases with semantic impairment show controlled retrieval deficits equivalent to PF+ cases or conceptual degradation similar to patients with SD. Independent of lesion location, the SA subgroups showed similarities, whereas SD patients showed a qualitatively distinct semantic impairment. Relative to SD, both TPC and PF+ SA subgroups: (1) showed few correlations in performance across tasks with differing control demands, but a strong relationship between tasks of similar difficulty; (2) exhibited attenuated effects of lexical frequency and concept familiarity, (3) showed evidence of poor semantic regulation in their verbal output - performance on picture naming was substantially improved when provided with a phonological cue, and (4) showed effects of control demands, such as retrieval difficulty, which were equivalent in severity across TPC and PF+ groups. These findings show that semantic impairment in SA is underpinned by damage to a distributed semantic control network, instantiated across anterior and posterior cortical areas.


Risk of malnutrition is associated with mental health symptoms in community living elderly men and women: the Tromsø study.
Little research has been done on the relationship between malnutrition and mental health in community living elderly individuals. In the present study, we aimed to assess the associations between mental health (particularly anxiety and depression) and both the risk of malnutrition and body mass index (BMI, kg/m(2)) in a large sample of elderly men and women from Tromsø, Norway. In a cross-sectional survey, with 1558 men and 1553 women aged 65 to 87 years, the risk of malnutrition was assessed by the Malnutrition Universal Screening Tool ('MUST'), and mental health was measured by the Symptoms Check List 10 (SCL-10). BMI was categorised into six groups (< 20.0, 20.0-22.4, 22.5-24.9, 25.0-27.4, 27.5-29.9, ≥ 30.0 kg/m(2)). The risk of malnutrition (combining medium and high risk) was found in 5.6% of the men and 8.6% of the women. Significant mental health symptoms were reported by 3.9% of the men and 9.1% of the women. In a model adjusted for age, marital status, smoking and education, significant mental health symptoms (SCL-10 score ≥ 1.85) were positively associated with the risk of malnutrition (odds ratio 3.9 [95% CI 1.7-8.6] in men and 2.5 [95%CI 1.3-4.9] in women), the association was positive also for subthreshold mental health symptoms. For individuals with BMI < 20.0 the adjusted odds ratio for significant mental health symptoms was 2.0 [95% CI 1.0-4.0]. Impaired mental health was strongly associated with the risk of malnutrition in community living elderly men and women and this association was also significant for subthreshold mental health symptoms.


RAS pathways in prostate cancer - mediators of hormone resistance?
Although not frequently mutated in prostate cancer Ras isoforms play a pivotal role in multiple pathways that have been implicated in prostate cancer progression to androgen independence. These have included growth factor and cytokine induced activation of the androgen receptor and its coregulators by post translational modification. Current evidence suggests that Ras is also required for androgen receptor activation in hormone sensitive cells. More recently Ras has been shown to work synergistically with other pathways to promote prostate tumorigenesis. We review the multiple lines of evidence implicating Ras as therapeutic target in androgen dependent and independent prostate cancer.


c-Fos-induced activation of a TATA-box-containing promoter involves direct contact with TATA-box-binding protein.
Transcriptional activation in eukaryotes involves protein-protein interactions between regulatory transcription factors and components of the basal transcription machinery. Here we show that c-Fos, but not a related protein, Fra-1, can bind the TATA-box-binding protein (TBP) both in vitro and in vivo and that c-Fos can also interact with the transcription factor IID complex. High-affinity binding to TBP requires c-Fos activation modules which cooperate to activate transcription. One of these activation modules contains a TBP-binding motif (TBM) which was identified through its homology to TBP-binding viral activators. This motif is required for transcriptional activation, as well as TBP binding. Domain swap experiments indicate that a domain containing the TBM can confer TBP binding on Fra-1 both in vitro and in vivo. In vivo activation experiments indicate that a GAL4-Fos fusion can activate a promoter bearing a GAL4 site linked to a TATA box but that this activity does not occur at high concentrations of GAL4-Fos. This inhibition (squelching) of c-Fos activity is relieved by the presence of excess TBP, indicating that TBP is a direct functional target of c-Fos. Removing the TBM from c-Fos severely abrogates activation of a promoter containing a TATA box but does not affect activation of a promoter driven only by an initiator element. Collectively, these results suggest that c-Fos is able to activate via two distinct mechanisms, only one of which requires contact with TBP. Since TBP binding is not exhibited by Fra-1, TBP-mediated activation may be one characteristic that discriminates the function of Fos-related proteins.


Identification of cell wall-associated proteins from Phytophthora ramorum.
The oomycete genus Phytophthora comprises a large group of fungal-like plant pathogens. Two Phytophthora genomes recently have been sequenced; one of them is the genome of Phytophthora ramorum, the causal agent of sudden oak death. During plant infection, extracellular proteins, either soluble secreted proteins or proteins associated with the cell wall, play important roles in the interaction with host plants. Cell walls of P. ramorum contain 1 to 1.5% proteins, the remainder almost exclusively being accounted for by glucan polymers. Here, we present an inventory of cell-wall-associated proteins based on mass spectrometric sequence analysis of tryptic peptides obtained by proteolytic digestion of sodium dodecyl sulfate-treated mycelial cell walls. In total, 17 proteins were identified, all of which are authentic secretory proteins. Functional classification based on homology searches revealed six putative mucins or mucin-like proteins, five putative glycoside hydrolases, two transglutaminases, one annexin-like protein, the elicitin protein RAM5, one protein of unknown function, and one Kazal-type protease inhibitor. We propose that the cell wall proteins thus identified are important for pathogenicity.


Regular dark chocolate consumption's reduction of oxidative stress and increase of free-fatty-acid mobilization in response to prolonged cycling.
This study investigated the effects of regular consumption of dark chocolate (DC), rich in cocoa polyphenols, on plasma metabolites, hormones, and markers of oxidative stress after prolonged exhaustive exercise. Twenty active men cycled at 60% maximal oxygen uptake (VO2max) for 1.5 hr, with the intensity increased to 90% VO2max for a 30-s period every 10 min, followed by a ride to exhaustion at 90% VO2max. In the 2 wk before exercise participants consumed 40 g of DC or an isocarbohydrate-fat control cocoa liquor-free chocolate (CON) twice daily and once 2 hr before exercise in a randomized, counterbalanced, crossover design. Venous blood samples were taken immediately before exercise, postexercise (fixed duration), postexhaustion, and after 1 hr of recovery. F2-isoprostanes were significantly lower (post hoc tests: p < .001) at exhaustion and after 1 hr of recovery with DC. Oxidized low-density lipoproteins were significantly lower with DC (p < .001) both before and after exercise and at exhaustion. DC was also associated with ~21% greater rises in free fatty acids during exercise (main effect: p < .05). Changes in circulating glucose, insulin, glucagon, cortisol, and interleukin (IL)-6, IL-10, and IL-1ra were unaffected by treatment. Time to exhaustion at 90% VO2max was not significantly different between trials (398 ± 204 and 374 ± 194 s for DC and CON, respectively). These results suggest that regular DC intake is associated with reduced oxidative-stress markers and increased mobilization of free fatty acids after exercise but has no observed effect on exercise performance.


Perceiving distance: a role of effort and intent.
Perceiving egocentric distance is not only a function of the optical variables to which it relates, but also a function of people's current physiological potential to perform intended actions. In a set of experiments, we showed that, as the effort associated with walking increases, perceived distance increases if the perceiver intends to walk the extent, but not if the perceiver intends to throw. Conversely, as the effort associated with throwing increases, perceived distance increases if people intend to throw to the target, but not if they intend to walk. Perceiving distance combines the geometry of the world with our behavior goals and the potential of our body to achieve these goals.


Nutritional effects on the growth and fatness of broilers.
1. Broiler chickens given diets high in protein, or choice-fed on a high protein balancer, had much lower abdominal fat contents than those reported in many recent experiments. The values for males were 10.8 g/kg liveweight at 56 d at 2.43 kg liveweight in one experiment in Scotland and 16.0 g/kg liveweight at 42 d at 1.93 kg liveweight in another in South Africa. For females the values were 18.8 g/kg liveweight at 56 d at 2.15 kg liveweight in Scotland and 15.7 g/kg liveweight at 42 d at 1.60 kg in South Africa. 2. The content of abdominal fat was, in general, increased by reducing the protein content of the diet or by dilution of the food with oil or starch. It was, in general, reduced by diluting the food with dietary fibre which also reduced liveweight gain. 3. The results are consistent with the idea that chickens attempt to control their food intake so that they achieve a particular fatness. This level of fatness differs between the sexes and between degrees of maturity.


Usefulness of Measuring Changes in SOFA Score for the Prediction of 28-Day Mortality in Patients With Sepsis-Associated Disseminated Intravascular Coagulation.
The primary end point for sepsis trial is 28-day mortality. However, additional methods for determining the efficacy may have benefits. The purpose of this study was to search a useful indicator of anticoagulant therapy in patients with sepsis with disseminated intravascular coagulation (DIC). Data from 323 patients with sepsis with coagulopathy treated with antithrombin supplementation were analyzed. The changes in the Sequential Organ Failure Assessment (Δ SOFA) score, the overt-DIC (Δ overt-DIC) score, and the Japanese Society for Acute Medicine DIC (Δ JAAM DIC) score from baseline to day 7 were retrospectively analyzed in relation to the 28-day mortality. Significant correlations were found between the 28-day mortality and Δ SOFA, Δ overt-DIC score, and Δ JAAM DIC score. The accuracy of the prediction was higher for Δ SOFA (80.5%) than for Δ overt-DIC (66.7%, P < .001). The areas under the curve for mortality calculated using a receiver operating characteristic curve analysis were 0.812 for Δ SOFA, 0.655 for Δ overt-DIC, and 0.693 for Δ JAAM DIC. The mortality rate was significantly lower among cases with an improved SOFA score compared to those without an improvement. The Δ SOFA had the strongest association with the 28-day mortality in patients with sepsis and DIC.


Is grip strength associated with length of stay in hospitalised older patients admitted for rehabilitation? Findings from the Southampton grip strength study.
identification of patients at risk of prolonged hospital stay allows staff to target interventions, provide informed prognosis and manage healthcare resources. Admission grip strength is associated with discharge outcomes in acute hospital settings. to explore the relationship between grip strength and length of stay in older rehabilitation in-patients. single-centre prospective cohort study. community hospital rehabilitation ward. one hundred and ten patients aged 70 years and over. data on age, height, weight, body mass index (BMI), co-morbidities, medication, residence, grip strength, physical function, cognitive function, frailty, falls, discharge destination and length of stay were recorded. higher grip strength was associated with reduced length of stay, characterised by an increased likelihood of discharge to usual residence among male rehabilitation in-patients (hazard ratio 1.09 (95% confidence interval 1.01, 1.17) per kilo increase in grip strength, P = 0.02) after adjustment for age and size. this is the first prospective study to show that stronger grip strength, particularly among male in-patients, is associated with a shorter length of stay in a rehabilitation ward. This is important because it demonstrates that grip strength can be discriminatory among frailer people. Further research into the clinical applications of grip strength measurement in rehabilitation settings is needed.


Talking it over with the gals: an analysis of editorials by Phyllis Stern.
Editorials written by a journal editor proved a glimpse of that individual's philosophy and opinions about topical events. The purpose of the present project was to analyze the editorials of Phyllis Noerager Stern during the years that she served as editor-in-chief of Health Care for Women International (1983-2001). Using ethnographic content analysis, 65 editorials were examined to ascertain: (1) what event triggered the editorial; (2) what aim or objective could be identified; and (3) what rhetorical devices the editor employed. Personal narratives and editorials about the abuse/exploitation of women made up the largest categories. Stern's grounded theory orientation, fascination with other cultures, and zest for life were evident throughout the corpus of editorials.


Medical ethics education: coming of age.
Medical ethics education is instruction that endeavors to teach the examination of the role of values in the doctor's relationship with patients, colleagues, and society. It is one front of a broad curricular effort to develop physicians' values, social perspectives, and interpersonal skills for the practice of medicine. The authors define medical ethics education as more clinically centered than human values education and more inclusive of philosophical, social, and legal issues than is interpersonal skills training. The authors review the history of the emergence of medical ethics education over the last 20 years, examine the areas of consensus that have emerged concerning the general objectives and premises for designing medical ethics programs, and describe teaching objectives and methods, course content, and program evaluation used in such programs on both preclinical and clinical levels. The four interrelated requirements for successful institutionalization of medical ethics education programs are defined and discussed, and the paper ends with an overview of the uncertain future of medical ethics education, an accepted but still not fully mature part of physician training in the United States. An extensive reference list accompanies the article.


Saliva in relation to dental erosion before and after radiotherapy.
Low saliva flow and abnormal saliva composition are common conditions after radiotherapy for oral cavity and pharyngeal cancer. Both conditions increase the susceptibility to dental caries and erosion, which may be further accelerated by changes in food preferences. The aim of this study was to determine changes in saliva flow and susceptibility to erosive challenges in pharyngeal cancer patients before and after radiotherapy to the head and neck. The erosive potential of sucking acidic candies with and without calcium was determined in nine patients (50-68 years) before and after receiving a radiation dose of 66 Gy to the head and neck area. The erosive potential was evaluated from saliva degree of saturation with respect to hydroxyapatite (HAp) and by dissolution of HAp in candy-stimulated saliva. Sucking acidic candies increased saliva flow rates ≈ 17-fold before as well as after radiotherapy (p < 0.001). However, significantly lower unstimulated (p < 0.05) and stimulated (p < 0.01) saliva flow rates were obtained after radiotherapy. Also, saliva became more under-saturated with respect to HAp during (p < 0.01) and in a period after sucking the candies (p < 0.01). HAp dissolution was significantly lower with the candy containing calcium compared with the control candy, both before and after radiotherapy (p < 0.001 and p < 0.05). Radiotherapy to the head and neck area significantly reduced saliva flow and altered saliva composition in a way that may increase the susceptibility to dental disease. However, saliva could be stimulated by acidic candies, which could be made nearly non-erosive even in irradiated patients.


Combined effects of brain-derived neurotrophic factor immobilized poly-lactic-co-glycolic acid membrane with human adipose-derived stem cells and basic fibroblast growth factor hydrogel on recovery of erectile dysfunction.
Erectile dysfunction (ED) is the most frequent long-term problem after radical prostatectomy. We aimed to evaluate whether the use of combination therapy with basic fibroblast growth factor (bFGF)-hydrogel on corpus cavernosum and with adipose-derived stem cells (ADSCs) and brain-derived neurotrophic factor (BDNF)-immobilized poly-lactic-co-glycolic acid (PLGA) membrane on the cavernous nerve (CN) could improve erectile function in a rat model of bilateral cavernous nerve crush injury (BCNI). Rats were randomly divided into five groups (n=15 per group): a normal group (N group), a group receiving saline application after bilateral cavernous nerve crush injury (BCNI), a group undergoing bFGF-hydrogel injection in the corpus cavernosum after BCNI (bFGF), a group receiving ADSC application covered with BDNF-membrane after BCNI (ADSC/BDNF), and a group undergoing coadministration of bFGF-hydrogel injection and BDNF-membrane with ADSCs after BDNF (bFGF+ADSC/BDNF). Four weeks postoperatively, the erectile function was assessed by detecting the ratio of intracavernous pressure (ICP) to mean arterial pressure (MAP). Smooth muscle and collagen contents were measured using Masson's trichrome staining. Neuronal nitric oxide synthase (nNOS) expression in the dorsal penile nerve was detected by immunostaining. The protein expression of the α-smooth muscle actin (α-SMA) and the cyclic guanosine monophosphate (cGMP) level of the corpus cavernosum were quantified by western blot and cGMP assay, respectively. In the bFGF+ADSC/BDNF group, the erectile function was significantly elevated compared with the BCNI and other treated groups and showed a significantly increased smooth muscle/collagen ratio, nNOS content, α-SMA expression, and cGMP level. In particular, there were no statistical differences in the ICP/MAP ratio, smooth muscle/collagen ratio, and α-SMA and cGMP levels between the bFGF+ADSC/BDNF group and normal group. Application of the BDNF-immobilized PLGA membrane with human ADSC into the CN and bFGF-incorporated hydrogel into the corpus carvernosum improved nearly normal erectile function in a rat model of postprostatectomy ED. This result suggests that a combined application of bFGF+ADSC/BDNF might be a promising treatment for postprostatectomy ED.


International differences in treatment effect: do they really exist and why?
With the increasing globalization of clinical trials, the opportunity exists to explore potential geographic differences in treatment effect within any major trial. Such geographic differences may arise because of international differences in patient selection, medical practice, or evaluation of outcomes, and such international variations need better documentation in trial reports. Appropriate pre-defined statistical analyses, including statistical tests of interaction regarding geographic heterogeneity in treatment effect, are important. Geographic variations are a particularly tricky form of subgroup analysis: they lack statistical power, are at best hypothesis-generating and can generate more confusion than insight. Referring to key examples, e.g. the PLATO and MERIT-HF, we emphasize the need for caution in interpreting evidence of potential geographic inconsistencies in treatment effect. Although it is appropriate to explore any biological or practical reasons for apparent geographic anomalies in treatment effect, the play of chance is often the most plausible and wise interpretation.


Evaluation of the sealing ability of different root canal sealers: a combined SEM and micro-CT study.
The purpose of this study was to analyze the ability of multiple compounds to seal the dental tubules using scanning electron microscopy (SEM) and micro-computed tomogra-phy (micro-CT). Twenty-four single-root human mandibular premolars were selected and instrumented with nickel-titanium rotary file and the final file size was #40/06. They were then randomly allocated into 2 groups, and all samples were filled with single cone gutta-percha (#40/06) and one of the tested sealers (AH Plus and EndoSequence BC sealers). All specimens were scanned using micro-CT and then three from each group were randomly selected for SEM analysis. According to SEM, both root canal sealers showed sufficient adaptation to dentin along the whole length of the root canal, though the coronal sections presented superior sealing than the apical sections. Micro porosity analyses revealed that the volume of closed pores and the surface of closed pores had the largest values in the coronal sections, followed by the middle and the apical sections for both sealants (p<0.05). However, no significant difference was observed for those two parameters between AH Plus and EndoSequence BC sealers in any of the three sections (p>0.05), whereas they were larger in the apical section when the AH Plus sealer was used. By using the single cone technique, neither EndoSequence or AH Plus pro-vides a porosity-free root canal filling. The EndoSequence BC sealer may have similar sealing abilities regarding the whole root canal as the AH Plus sealer. A better sealing effect could be obtained in the coronal and middle sections of a root canal than the apical part by using the tested sealers.


Comparing efficiency of micro-RNA and mRNA biomarker liberation with microbubble-enhanced ultrasound exposure.
Blood biomarkers are potentially powerful diagnostic tools that are limited clinically by low concentrations, the inability to determine biomarker origin and unknown patient baseline. Recently, ultrasound has been shown to liberate proteins and large mRNA biomarkers, overcoming many of these limitations. We have since demonstrated that adding lipid-stabilized microbubbles elevates mRNA concentration an order of magnitude compared with ultrasound without microbubbles, in vitro. Unfortunately the large size of some mRNA molecules may limit efficiency of release and hinder efficacy as an ultrasound-liberated biomarker. We hypothesize that smaller molecules will be released more efficiently with ultrasound than larger molecules. Although investigation of large libraries of biomarkers should be performed to fully validate this hypothesis, we focus on a small subset of mRNA and micro-RNAs. Specifically, we focus on miR-21 (22 base pairs [bp]), which is upregulated in certain forms of cancer, compared with previously investigated mammaglobin mRNA (502 bp). We also report release of micro-RNA miR-155 (22 bp) and housekeeping rRNA S18 (1869 bp). More than 10 million additional miR-21 copies per 100,000 cells are released with ultrasound-microbubble exposure. The low- molecular-weight miR-21 proved to be liberated 50 times more efficiently than high-molecular-weight mammaglobin mRNA, releasing orders of magnitude more miR-21 than mammaglobin mRNA under comparable conditions.