Defining emergency department episodes by severity and intensity: A 15-year study of Medicare beneficiaries.
Episodes of Emergency Department (ED) service use among older adults previously have not been constructed, or evaluated as multi-dimensional phenomena. In this study, we constructed episodes of ED service use among a cohort of older adults over a 15-year observation period, measured the episodes by severity and intensity, and compared these measures in predicting subsequent hospitalization. We conducted a secondary analysis of the prospective cohort study entitled the Survey on Assets and Health Dynamics among the Oldest Old (AHEAD). Baseline (1993) data on 5,511 self-respondents >or=70 years old were linked to their Medicare claims for 1991-2005. Claims then were organized into episodes of ED care according to Medicare guidelines. The severity of ED episodes was measured with a modified-NYU algorithm using ICD9-CM diagnoses, and the intensity of the episodes was measured using CPT codes. Measures were evaluated against subsequent hospitalization to estimate comparative predictive validity. Over 15 years, three-fourths (4,171) of the 5,511 AHEAD participants had at least 1 ED episode, with a mean of 4.5 episodes. Cross-classification indicated the modified-NYU severity measure and the CPT-based intensity measure captured different aspects of ED episodes (kappa = 0.18). While both measures were significant independent predictors of hospital admission from ED episodes, the CPT measure had substantially higher predictive validity than the modified-NYU measure (AORs 5.70 vs. 3.31; p < .001). We demonstrated an innovative approach for how claims data can be used to construct episodes of ED care among a sample of older adults. We also determined that the modified-NYU measure of severity and the CPT measure of intensity tap different aspects of ED episodes, and that both measures were predictive of subsequent hospitalization.


Culture, Religion and the State: Towards a Multidisciplinary Approach to Ensuring Public Health During the COVID-19 Pandemic (and Beyond).
This research investigates different approaches by cultural, religious and political groups in ensuring public health during the COVID-19 pandemic. During the pandemic, several methods were done by governments and healthcare institutions to mitigate the spread of the virus. This study uses content analysis, specifically a literature review, on existing online literature found in online journals, books, and reports relevant to the COVID-19 pandemic. The paper presents the available qualitative data including substantive findings as well as theoretical and methodological foundations relevant to the topic. Culture, religion and the state have roles to play in ensuring public health during the COVID-19 pandemic. These three areas can help in mitigating the transmission of the virus, managing patients, and aiding the rollout of COVID-19 vaccines. This paper ends by suggesting that, in ensuring public health during the COVID-19 pandemic, a multidisciplinary approach, involving a concerted effort among culture, religion and the state, is necessary. This approach is also recommended for any public health crisis in the future.


[Hyperkinetic syndrome and diet--a therapeutic alliance? (author's transl)].
The international literature on dietary treatment of the hyperkinetic syndrome is reported and analyzed critically. So far there is no sufficient scientific evidence in favour of this therapeutic intervention with hyperkinetic children.


Characterization of the stemness potency of mammospheres isolated from the breast cancer cell lines.
Stemness phenotype mammospheres established from cell lines and tissues taken from autopsy can be used to test and to identify the most sensitive drugs for chemotherapy. Therefore, the aim of the present study was isolation and characterization of cancer stem cells derived from MCF7, MDA-MB231, and SKBR3 breast cancer cell lines to demonstrate the stemness phenotypes of mammospheres generated for further their applications in therapeutic approaches. In this study, two luminal subtypes of cell lines, MCF7 and SKBR3 and a basal subtype cell line, MDA-MB-231, were chosen. Mammosphere culturing was implemented for breast cancer stem cells isolation and mammosphere formation efficiency. At the next step, CD44+/CD24- cell ratio, Oct4 and Nanog mRNA levels, proliferation rate, migration rate of mammospheres, and drug resistance (in third passage) were evaluated. In addition, tumorigenicity of mammospheres in the chick embryo model was evaluated and compared through the chick chorioallantoic membrane assay. Among mammospheres formed in all three cell lines, MCF7 had the highest mammosphere formation efficiency. CD24 marker (a differentiation marker for the breast cancer cells) was significantly reduced in the mammospheres generated from MCF7 and SKBR3, during three passages. Also, Oct4 and Nanog transcript levels were significantly higher in all three types of mammospheres, as compared with their cell lines. Proliferation, migration rate, and drug resistance of mammospheres generated from all three cell lines were found to be significantly higher. Tumorigenicity of MCF7 mammospheres was confirmed through tumor size measurement. Also, tumorigenicity of MCF7 and SKBR3 mammospheres was confirmed through more migration from ectoderm to mesoderm and endoderm. We succeeded to establish the technology that can be extended to tissue in the future. We have demonstrated a number of mammospheres can be generated from cell lines. Also, cells with different molecular features showed different stemness phenotypes.


Gender differences in health related quality of life among people living with HIV on highly active antiretroviral therapy in Mekelle Town, Northern Ethiopia.
Health related quality of life (HRQOL) is an important outcome measure for highly active antiretroviral treatment program. In Ethiopia, studies revealed that there are improved qualities of life among adults living with the viruses taking antiretroviral therapy but there is no explicit data showing gender differences in health related quality of life. Aim. To assess gender differences in HRQOL and its associated factors among people living with HIV and on highly active antiretroviral therapy in public health institutions of Mekelle town, Northern Ethiopia. A comparative cross-sectional study was conducted among 494 adult people living with HIV taking ART services. Quality of life was measured using WHOQOL-HIV BREF. There was a statistically significant gender difference (P < 0.05) in HRQOL among PLHIV on HAART. Females had low score in all HRQOL domains. High perceived stigma was strongly associated with poor psychological quality of domain among both female and male groups with [AOR = 2.89(1.69,4.96)] and [AOR = 2.5(1.4,4.4)], respectively. There was statistically significant gender difference in all quality of life domains. Public health interventions to improve HRQOL of PLHIV should take in to account the physical, psychological, social, environmental, and spiritual health of PLHIV during treatment, care, and support.


Ovarian fibrothecoma with massive edema.
We report a rare case of ovarian fibrothecoma with massive edema. The patient was a 59-year-old woman with a left ovarian mass measuring 11 x 10 x 7 cm. Magnetic resonance images revealed a solid mass showing unhomogeneous content with predominantly high signal intensity on T2-weighted image. Microscopically, the ovarian mass was composed of a cellular area and an edematous hypocellular area. The latter accounted for more than 75% of the tumor. In the cellular area, spindle-shaped or plump tumor cells were randomly distributed or arranged in a fascicular fashion. These cells contained abundant intracytoplasmic lipid. There was dense collagenous connective tissue in the stroma of the cellular areas. In contrast, in the edematous areas spindle or stellate cells were scattered. Alcian blue stain revealed only a small amount of stromal mucin even in the edematous areas. The microscopic findings were consistent with that of fibrothecoma with massive edema. The present case must be differentiated from massive edema of the ovary and sclerosing stromal tumor of the ovary. Immunohistochemistry was not helpful in distinguishing them. The age of the patient and careful histologic observation are important.


[Electronic medical records: medical and legal aspects, privacy, safety, and legal validity].
Medical records must collect all data concerning in-hospital management of patients: data have to be verified and easily retrievable. Clinicians are responsible for both format and content of medical records. Respect of patient's privacy must be made sure both during on-line management and long-term storage of records. Computerization can offer many advantages to clinicians, but needs some significant adjustments: training and motivation of operators, arrangement of clinical processes and of administrative rules to technological developments. Nevertheless, some important results can be afforded: standardization of procedures, distribution of univocal, verified and ubiquitous data to all concerned operators, protection against undesired retrieval, reliability of effective reports. Preliminary condition is a clinical local area network, widespread into the institution. Database implementation must follow well accepted methodology: flow chart design of data dictionary, standardization of data coding, input of verified data, effective reporting. Access to data must be controlled by sophisticated and sure password system. Back-up of data must be automatically available with adequate timing and methodology. Respect of rules on patient's privacy must be realized whenever possible. Complex clinical records should be made available, containing data, signals and images (both single frames and dynamic sequences), due to continuous technical progress of diagnostic tools. Medical records must be available for long periods of time: database engine and managing tools must be selected among well accepted and largely available producers; informatic assistance must be assured for management and evolution of systems over the years.


[Immune response produced by anti-rabies vaccines in marmosets (Callithrix sp)].
In order to evaluate the immune response produced by rabies vaccines in new world nonhuman primates, thirty marmosets (Callithrix sp) were divided into five groups of six individuals and submitted to five different antirabies vaccination schemes using two distinct commercially available animal vaccines. The first was produced in suckling mouse brain (Fuenzalida and Palacios), and the second in NIL-2 cell culture. Post-vaccine serological monitoring was carried out periodically. The results showed that the Fuenzalida and Palacios vaccine was not able to protect the animals when using a single dose or even with a booster. But when submitted to a vaccination routine similar to that used for humans, the marmosets showed detectable antibodies, and only one succumbed to rabies after being challenged. In addition, the vaccine produced in NIL-2 cell culture induced high antibody levels in all vaccinated animals and all animals survived the viral challenge.


Tuberculosis infection in patients with rheumatoid arthritis and the effect of infliximab therapy.
According to the Centers for Disease Control and Prevention, the 1999 and 2000 incidence rates for tuberculosis (TB) in the US population were 6.4 and 5.8, respectively, per 100,000 persons. Recently, reports of TB following infliximab administration have raised questions regarding the rate of TB in patients with rheumatoid arthritis (RA) generally and in those treated with infliximab in clinical practice. We undertook this study to determine the baseline rate of TB in RA prior to the introduction of infliximab and to determine the rate of TB among those currently receiving infliximab. We surveyed patients with questionnaires, followed by detailed validation from medical records and physician reports. In study 1, we evaluated 10,782 RA patients in 1998-1999 prior to the widespread use of infliximab. In study 2, we evaluated 6,460 infliximab-treated patients in 2000-2002. In study 1, the lifetime rate of TB was 696 per 100,000 patients (95% confidence interval [95% CI] 547-872). Of these cases, 76.8% occurred prior to the onset of RA. During the period of prospective followup, 1 case of TB developed during 16,173 patient-years of followup, yielding a rate of 6.2 cases (95% CI 1.6-34.4) per 100,000 patients. In study 2, the TB incidence rate among infliximab-treated patients was 52.5 cases (95% CI 14.3-134.4) per 100,000 patient-years of exposure. Three of the 4 cases occurred in patients with a history of TB exposure, and no cases occurred in persons with recent TB skin tests or prophylaxis. The rate of TB is not increased in RA patients generally. Among infliximab-treated patients, the rate is 52.5 cases (95% CI 14.3-134.4) per 100,000 patient-years of exposure. A thorough medical history regarding TB, as well as tuberculin testing and radiographic examination (if indicated), should be an essential component of anti-tumor necrosis factor therapy.


rpoB sequence analysis as a novel basis for bacterial identification.
Comparison of the sequences of conserved genes, most commonly those encoding 16S rRNA, is used for bacterial genotypic identification. Among some taxa, such as the Enterobacteriaceae, variation within this gene does not allow confident species identification. We investigated the usefulness of RNA polymerase beta-subunit encoding gene (rpoB) sequences as an alternative tool for universal bacterial genotypic identification. We generated a database of partial rpoB for 14 Enterobacteriaceae species and then assessed the intra- and interspecies divergence between the rpoB and the 16S rRNA genes by pairwise comparisons. We found that levels of divergence between the rpoB sequences of different strains were markedly higher than those between their 16S rRNA genes. This higher discriminatory power was further confirmed by assigning 20 blindly selected clinical isolates to the correct enteric species on the basis of rpoB sequence comparison. Comparison of rpoB sequences from Enterobacteriaceae was also used as the basis for their phylogenetic analysis and demonstrated the genus Klebsiella to be polyphyletic. The trees obtained with rpoB were more compatible with the currently accepted classification of Enterobacteriaceae than those obtained with 16S rRNA. These data indicate that rpoB is a powerful identification tool, which may be useful for universal bacterial identification.


Eyes on CHARGE syndrome: Roles of CHD7 in ocular development.
The development of the vertebrate visual system involves complex morphogenetic interactions of cells derived from multiple embryonic lineages. Disruptions in this process are associated with structural birth defects such as microphthalmia, anophthalmia, and coloboma (collectively referred to as MAC), and inherited retinal degenerative diseases such as retinitis pigmentosa and allied dystrophies. MAC and retinal degeneration are also observed in systemic congenital malformation syndromes. One important example is CHARGE syndrome, a genetic disorder characterized by coloboma, heart defects, choanal atresia, growth retardation, genital abnormalities, and ear abnormalities. Mutations in the gene encoding Chromodomain helicase DNA binding protein 7 (CHD7) cause the majority of CHARGE syndrome cases. However, the pathogenetic mechanisms that connect loss of CHD7 to the ocular complications observed in CHARGE syndrome have not been identified. In this review, we provide a general overview of ocular development and congenital disorders affecting the eye. This is followed by a comprehensive description of CHARGE syndrome, including discussion of the spectrum of ocular defects that have been described in this disorder. In addition, we discuss the current knowledge of CHD7 function and focus on its contributions to the development of ocular structures. Finally, we discuss outstanding gaps in our knowledge of the role of CHD7 in eye formation, and propose avenues of investigation to further our understanding of how CHD7 activity regulates ocular and retinal development.


Antibacterial effect against both Gram-positive and Gram-negative bacteria via lysozyme imprinted cryogel membranes.
The development of novel biocompatible and cost effective cryogel membrane which shows enhanced antimicrobial properties in order to use for several approaches such as wound dressing, scaffold or food packaging was aimed in this study. A super macro porous lysozyme imprinted cryogel membranes showing antibacterial effect against both Gram-positive and Gram-negative bacteria were prepared by using molecular imprinting technique. N-methacryloyl-(L)-histidine methyl ester (MAH) was used as the pseudo specific ligand and complexed with Cu++ in order to provide metal ion coordination between MAH and template molecule (lysozyme). Comparing the antibacterial activity of different lysozyme concentrations, cryogel membranes were prepared in three different concentrations. To synthesize Poly (hydroxyethyl methacrylate-N-methacryloyl-(L)-histidine methylester) P(HEMA-MAH) cryogel membrane, free radical polymerization initiated by N, N, N', N'-tetramethylene diamine (TEMED) and ammonium persulfate (APS) was carried out at -12 °C. The characterization of the lysozyme imprinted cryogel membrane was accomplished by using scanning electron microscopy (SEM), swelling degree measurements and Fourier transform infrared spectroscopy-attenuated total reflectance (FTIR-ATR) spectroscopy. The cytotoxicity test of produced membrane was performed by using mouse fibroblast cell line L929. The antibacterial activity of P(HEMA-MAH) lysozyme molecular imprinted [P(HEMA-MAH) Lyz-MIP] cryogel membranes against Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) were determined by Kirby-Bauer membranes diffusion and viable cell counting methods. When the antibacterial effect of P(HEMA-MAH) Lyz-MIP cryogel membranes were evaluated, it was found that P(HEMA-MAH) Lyz-MIP cryogel membranes had stronger antibacterial effects against Gram-negative E. coli bacteria even in low lysozyme concentrations. In addition, 100% bacterial inhibition was detected for both of two bacteria at increasing lysozyme concentrations.


Soil mixing to decrease surface stratification of phosphorus in manured soils.
Continual applications of fertilizer and manure to permanent grassland or no-till soils can lead to an accumulation of P at the surface, which in turn increases the potential for P loss in overland flow. To investigate the feasibility of redistributing surface stratified P within the soil profile by plowing, Mehlich-3 P rich surface soils (128-961 mg kg(-) in 0-5 cm) were incubated with lower-P subsoil (16-119 mg kg(-1) in 5-20 cm) for 18 manured soils from Oklahoma and Pennsylvania that had received long-term manure applications (60-150 kg P ha(-1) yr(-1) as dairy, poultry, or swine manure for up to 20 yr). After incubating a mixture of 5 g surface soil (0- to 5-cm depth) and 15 g subsoil (5- to 20-cm depth) for 28 d, Mehlich-3 P decreased 66 to 90% as a function of the weighted mean Mehlich-3 P of surface and subsoil (i.e.. 1:3 ratio) (r2 = 0.87). At Klingerstown, Northumberland County, south central Pennsylvania, a P-stratified Berks soil (Typic Dystrochrept) (495 mg kg(-1) Mehlich-3 P in 0- to 5-cm depth) was chisel plowed to about 25 cm and orchardgrass (Dactylis glomerata L.) planted. Once grass was established and erosion minimized (about 20 wk after plowing and planting), total P concentration in overland flow during a 30-min rainfall (6.5 cm h(-1)) was 1.79 mg L(-1) compared with 3.4 mg L(-1) before plowing, with dissolved P reduced from 2.9 to 0.3 mg L(-1). Plowing P-stratified soils has the potential to decrease P loss in overland flow, as long as plowing-induced erosion is minimized.


Nipple sparing mastectomy with prepectoral immediate prosthetic reconstruction without acellular dermal matrices: a single center experience.
Nipple-sparing mastectomy (NSM) with immediate prosthetic breast reconstruction (IPBR) is an oncologically accepted technique that allows to improve aesthetic results and patient quality of life. Traditionally, implant for reconstruction have been placed in a submuscolar (SM) plane, beneath the pectoralis major muscle (PMM). Recently, prepectoral (PP) placement of prosthesis is increasingly used in order to avoid morbidities related to manipulation of PMM. The aim of the present study was to report our experience with 209 NSMs and IPBR using a prepectoral approach and polyurethane-coated implant without acellular dermal matrices (ADMs). A retrospective review of breast cancer patients who underwent NSM followed by PP - IPBR from January 2018 to April 2021 was performed. Data were recorded in order to evaluate operative details, major complications and oncological outcomes. Aesthetic results and patient quality of life were measured by a specific "QOL assessment PRO" survey. Two hundred and nine patients (269 breasts) with PP - IPBR after NSM were included. Mean age was 47 (25-73) years and median follow-up was 14 (1-40) months. A simultaneous contralateral implant-based mammoplasty of symmetrization after unilateral NSM was carried out in six of 149 (4%) patients. Implant loss was observed in three of 209 patient (1.44%); two of 209 (0.96%) patients developed a full-thickness NAC necrosis that required excision. During follow-up one local relapse (0.48%) and two regional nodes recurrences (0,96%) was observed. Patient satisfaction, assessed using a personalized QOL Assessment PRO survey, in term of aesthetic results, chronic pain, shoulder dysfunction, sports activity, sexual and relationship life and skin sensibility, was excellent. Our experience shows that PP-IPBR using polyurethane-coated implant after NSM is a safe, reliable and effective alternative to traditional IPBR with excellent aesthetic outcomes and high patient quality of life; it is easy to perform, minimizes complications related to manipulation of PPM and reduces operative time while resulting also in a cost-effective technique.


Pseudomonas aeruginosa compared with Escherichia coli produces less endotoxemia but more cardiovascular dysfunction and mortality in a canine model of septic shock.
We investigated the effects of two different Gram-negative bacteria and radiation-induced leukopenia on endotoxemia, cardiovascular abnormalities, and mortality in a canine model of septic shock. Serial hemodynamics were measured in conscious dogs using radionuclide heart scans and thermodilution cardiac output catheters. Plasma endotoxin concentrations were determined with a chromogenic Limulus amebocyte lysate assay. Viable Pseudomonas aeruginosa or Escherichia coli implanted intraperitoneally produced concordant hemodynamic patterns of septic shock (p less than 0.01). Endotoxin concentrations were more than tenfold lower in dogs infected with P aeruginosa compared with E coli (p less than 0.0001). Despite lower endotoxin levels, P aeruginosa-infected dogs had a higher mortality (p less than 0.01), more severe hypotension (p less than 0.05), and greater depression of the left ventricular ejection fraction (p less than 0.05) than dogs with E coli sepsis. A nonlethal E coli challenge combined with leukopenia (induced by a nonlethal dose of radiation) resulted in a mortality of 60 percent (p less than 0.01) without greater cardiovascular dysfunction or higher endotoxin concentrations. These findings suggest that bacterial products other than endotoxin and host-related factors may be important contributors to the toxicity, cardiovascular instability, and mortality of Gram-negative septic shock. Quantitative determinations of plasma endotoxin are unlikely to correlate with the clinical severity of septicemia in heterogeneous patient populations infected with different Gram-negative organisms.


The Effect of an Overhead External Load Lifting and Lowering on Dynamic Postural Control in Subgroups of Low Back Pain.
Understanding postural control in low back pain (LBP) subgroups can help develop targeted interventions to improve postural control. The studies on this topic are limited. Therefore, the primary purpose of this study was to compare the postural control of LBP subgroups with healthy individuals during overhead load lifting and lowering. In this cross-sectional study, the participants were 52 with LBP and 20 healthy. The LBP patients were classified based on the O'Sullivan classification system into 21 flexion patterns and 31 active extension patterns. The participants lifted the box from their waists to their overheads and lowered it to their waists. Changes in postural control parameters were measured with a force plate system. The results of the analysis of variance showed that during load lifting, the mediolateral phase plane (p = .044) and the mean total velocity (p = .029) had significant differences between flexion patterns and healthy. Also, the load-lowering results showed that active extension patterns, compared with healthy, had significant differences in the anteroposterior-mediolateral phase plane (p = .042). The patients showed less postural sway than the healthy. The results in this work highlight the importance of identifying the homogenous subgroups in LBP and support the classification of heterogeneous LBP. Different subgroups exhibit different postural control behaviors. These behaviors can be due to the loading of various tissues during different tasks.


Investigation of a bifactor model of the Strengths and Difficulties Questionnaire.
The Strengths and Difficulties Questionnaire (SDQ) is used to measure psychopathological symptoms in children and adolescents from 4 to 17 years old, but its underlying structure is still a matter of debate. Indeed, on the basis of a systematic review of English and non-English articles conducted using multiple databases, 54 studies reporting on the factor structure of the SDQ were located. The original 5 first-order factor structure is generally supported by exploratory procedures, but support based on confirmatory factor analyses is not clear. We analysed data from 889 youths from the general French population, rated on the SDQ by their teachers. We tested the original model, hierarchical models and bifactor models. The best-fitting model is a bifactor model with the five a priori factors grouped in two global factors (Externalizing Disorders-Hyperactivity and Conduct-and Internalizing Disorders-Peer relationships and Emotions) and one Strength/Prosocial factor. However, we show that the Conduct-Specific factor should not be used in practice in its current state, that the Hyperactivity-Specific factor mainly covers hyperactivity rather than inattention, and that the Peer Problems-Specific factor mainly reflects a preference for solitude. Nevertheless, the measurement model proved to be fully invariant across gender and school levels (kindergarten, primary and secondary schools), with statistically significant differences in latent means between genders only. Beyond computing the five a priori scores when using the teacher ratings of the SDQ, our results prove the usefulness of computing Externalizing Disorders and Internalizing Disorders global scores.


Effects of monovalent cations, pH and temperature on the dissociation constant (KD) for the fluorescent indicator mag-fura-2 at different excitation wavelengths.
The use of fluorescent indicators to quantify intracellular magnesium ([Mg(2+)]i) requires accurate determination of the dissociation constant (K(D)). Ideally, the K(D) should be determined intracellularly; however, in practice this is difficult to achieve. An alternative method is to measure the K(D) in solutions which attempt to mimic the intracellular milieu. This study investigated the effect of monovalent cations, pH and temperature on the Mg(2+)-mag-fura-2 K(D) determined at individual excitation wavelength intensities of either 340 nm or 380 nm, with emission fluorescence set at 510 nm. Monovalent cation concentration had little effect on K(D). Temperatures in the range 25 degrees C to 37 degrees C had significant effects on K(D) measured at 340 nm and 380 nm. At 340 nm the K(D) values at 25 degrees C and 37 degrees C were 2.02 and 1.15 mM, respectively and at 380 nm 2.84 to 1.68 mM, respectively. Changes of pH in the range 6.5 to 8.5 had variable effects on the K(D) depending on the wavelength at which it was measured. The results illustrate not only how estimation of K(D) can vary with environmental conditions, including temperature and pH. but also the choice of wavelength.


Postoperative Myocardial Infarction in Administrative Data vs Clinical Registry: A Multi-Institutional Study.
Previous studies have documented significant differences between administrative data and registry data in the determination of postoperative MI. The goal of this study was to characterize discordance between administrative and registry data in the determination of postoperative myocardial infarction (MI). This study was performed using data from the American College of Surgeons NSQIP merged with administrative data from 8 different hospitals, between 2013 and 2015. From each of these sources, the occurrence of a postoperative MI, as ascertained by administrative data and NSQIP data, were compared. In each situation in which the 2 sources disagreed (discordance), a 2-clinician chart review was performed to generate a "gold standard" determination as to the occurrence of postoperative MI. A total of 43,289 operations met our inclusion criteria for analysis. Within this cohort a total of 230 cases of MI were identified by administrative data and/or NSQIP data (administrative rate 0.41%, NSQIP rate 0.42%). A total of 89 discordant ascertainments were identified, of which 42 were admin+/NSQIP- and 47 were admin-/NSQIP+. Accuracy (99.9% for both) and concordance (kappa = 0.89 [95% CI 0.86 to 0.92] for administrative data, kappa = 0.87 [95% CI 0.84 to 0.91] for NSQIP data) of the 2 systems were similar when compared against our gold standard (chart review). The majority of errors were related to false negatives, with sensitivity rates of 81% in both data sources. In this multi-institutional study, administrative data and NSQIP demonstrated a similar ability to determine the occurrence of postoperative MI. These findings do not demonstrate an advantage of registry data over administrative data in the determination of postoperative MI.


The SPA quartet: a family of WD-repeat proteins with a central role in suppression of photomorphogenesis in arabidopsis.
The Arabidopsis thaliana proteins suppressor of phytochrome A-105 1 (SPA1), SPA3, and SPA4 of the four-member SPA1 protein family have been shown to repress photomorphogenesis in light-grown seedlings. Here, we demonstrate that spa quadruple mutant seedlings with defects in SPA1, SPA2, SPA3, and SPA4 undergo strong constitutive photomorphogenesis in the dark. Consistent with this finding, adult spa quadruple mutants are extremely small and dwarfed. These extreme phenotypes are only observed when all SPA genes are mutated, indicating functional redundancy among SPA genes. Differential contributions of individual SPA genes were revealed by analysis of spa double and triple mutant genotypes. SPA1 and SPA2 predominate in dark-grown seedlings, whereas SPA3 and SPA4 prevalently regulate the elongation growth in adult plants. Further analysis of SPA2 function indicated that SPA2 is a potent repressor of photomorphogenesis only in the dark but not in the light. The SPA2 protein is constitutively nuclear localized in planta and can physically interact with the repressor COP1. Epistasis analysis between spa2 and cop1 mutations provides strong genetic support for a biological significance of a COP1-SPA2 interaction in the plant. Taken together, our results have identified a new family of proteins that is essential for suppression of photomorphogenesis in darkness.


Hypobaric hypoxia and villous trophoblast: evidence that human pregnancy at high altitude (3600 m) perturbs epithelial turnover and coagulation-fibrinolysis in the intervillous space.
Spatial relationships between fibrin-type fibrinoid and regions of villous trophoblast were examined in order to address two main questions: [1] is high-altitude pregnancy accompanied by changes in the sizes of trophoblast compartments (cytotrophoblast, syncytiotrophoblast, denudation sites)?, and [2] do highland placentae differ in the amounts and distribution patterns of perivillous fibrin-type fibrinoid? Placentae were collected from two ethnic groups completing term pregnancies at low (400 m above sea level; n=25) and high (3600 m; n=45) altitude in Bolivia. Masson trichrome-stained sections were sampled randomly and analysed stereologically to estimate compartment volumes and surfaces. Comparisons were drawn using variance, Chi-squared and contingency table analyses. At high altitude, birthweights were 265 g lower and placentas had a larger intervillous space (270 cf 181 cm(3)), less fibrin-type fibrinoid (4.1 cf 8.4 cm(3) by volume; 2570 cf 4430 cm(2) by surface area), less villous trophoblast (50 cf 73 cm(3)) and a smaller villous surface (5.6 cf 7.0 m(2)). Volumes were reduced in all syncytiotrophoblast compartments (with and without nuclear aggregations). Cytotrophoblast was maintained and its relative volume increased significantly (from 2.7 to 3.6 per cent of trophoblast volume). Decreases in villous surface area affected primarily thinner (nuclear aggregate-free) regions of syncytium. Regardless of altitude, fibrin-type fibrinoid was deposited non-randomly: it was preferentially located at sites of trophoblast denudation. Although no altitudinal differences in fibrin-type fibrinoid patterns were detected, absolute surfaces were diminished on denuded and thinner regions of trophoblast but not on syncytial knots or bridges. Ethnic differences at low altitude (relatively greater deposits on denudations in Amerindians) were minimized at high altitude. We conclude that pregnancy at high altitude alters the epithelial steady state (towards cytotrophoblast and away from syncytiotrophoblast) and the coagulation-fibrinolysis steady state in the intervillous space (to favour fibrinolysis over coagulation). Thinner regions of syncytiotrophoblast may be the main sites of greater fibrinolytic or anticoagulatory activity. The findings are partly consistent with results from in vitro studies which indicate that hypoxia stimulates proliferation of cytotrophoblast but impairs fusion into syncytium.


Social work in international health and medical assistance.
Welfare issues such as the poor, children, women, and the handicapped are dealt with in the field of development assistance. Few studies, however, have discussed development assistance from a social work point of view. This study analyzes the social work aspects of development assistance through a review of 60 health projects completed by the Japan International Cooperation Agency between 2000 and 2006. Although the term "social work" is ambiguous, several projects with diverse themes included what could be called social work. Projects conducted three types of activities: that for a target population of social works; that for the general population, which included its target population; and that not for a specific target population. Project interventions included both micro-level interventions and system development. There are several possible reasons why only a few projects included social work: 1) social work has a lower priority in development assistance than other areas such as health do, and 2) there are few relevant specialists who can handle a wide range of social work interventions. Donor agencies are gradually focusing more on social work aspects in their projects. Since social work will likely become a greater necessity in the field of development assistance for developing countries in the near future, donor nations and agencies will need to be prepared more adequately to respond to social work needs.


Reduced oncotic pressure after shock: a proposed mechanism.
Hemorrhagic shock plus resuscitation leads to expansion of both interstitial fluid space (IFS) and intracellular fluid (ICF). THe IFS expansion is thought to reflect reduced serum colloid oncotic pressure (COP) from capillary leak of serum albumin (SA). This hypothesis was analyzed in 138 injured patients who received an average of 13.6 units of blood and 10.7 L of saline solution for correction of shock. Plasma volume (PV), extracellular fluid (ECF) by inulin space technique, SA level, percent albumin leak (PAL), total intravascular albumin content (TIAC), COP, IFS, weight gain, and time of injury were analyzed. Patients studied during the postoperative fluid sequestration phase had a low to normal PV (2.9 L) with a high IFS (15.2 L) and a low PV-ECF ratio (0.15). Although the SA level (2.8 g/dL), TIAC (81 g), and COP (11.9 mOsm/L) were low, the PAL was also low (6.0%/hr; normal, 7%/hr). Patients studied during the fluid mobilization phase had a high PV (3.2 L) and normal PV-ECF ratio (0.21), whereas the SA level (3.0 g/dL), TIAC (103 g), and COP (14.1 mOsm/L) remained low. Despite these findings, the PAL was high (8.0%/hr). The fall in the PV-ECF ratio during the postresuscitation fluid sequestration phas is associated with but not due to reduced TIAC and COP, both of which remained low after the PV-ECF ratio has normalized during the mobilization phase. Contrary to prior reports, albumin leak is not responsible for reduced COP, TIAC, and postresuscitation weight gain. Presumably, the low COP results from decreased reentry of albumin into the plasma volume due to entrapment within the IFS matrix.


Can a specialist parasite species of a widespread and common host species be rare? The case of Spinitectus inermis (Nematoda: Cystidicolidae) in eels Anguilla anguilla.
The claim by many authors that Spinitectus inermis (Zeder, 1800), a narrowly specific parasite of European eels Anguilla anguilla (L.), is a rare species is considered at three levels: its geographical range, its frequency of occurrence compared to other eel parasites and its relative abundance in component communities. The parasite is widely distributed in freshwater throughout the European range of the eel but its occurrence is erratic and unpredictable, being known from only 8 countries. Surveys of eel parasites in the United Kingdom and in Continental Europe show that it is present in only 13% of British and 29% of continental localities. This satisfies one of the criteria for rarity. When present, its prevalence ranges from 1.8% to 43.3%, so it can be considered rare in some localities but in a few it may be common and on occasion it may be the dominant species in the gastro-intestinal community. Populations of S. inermis are almost always characterised by high levels of overdispersion, even at low prevalence. The species also displays an ability to colonise a locality following introduction there. Overall it meets many of the criteria of a rare species including a restricted distribution and a low frequency of occurrence and so it can be considered to exhibit diffusive rarity.


Pleurotus abieticola Polysaccharide Alleviates Hyperlipidemia Symptoms via Inhibition of Nuclear Factor-κB/Signal Transducer and Activator of Transcription 3-Mediated Inflammatory Responses.
Hyperlipidemia (HLP) is a metabolic syndrome induced by obesity, which has been widely recognized as a significant threat to human health. Pleurotus abieticola, an edible lignin-degrading fungus, remains relatively understudied in terms of its bioactivity and medicinal properties. In this study, the lipid-lowering effect of Pleurotus abieticola polysaccharide (PAPS1) was systematically explored in high-fat diet (HFD)-induced HLP mice. The findings demonstrated that the administration of PAPS1 significantly inhibited bodyweight gain, ameliorated blood glucose and lipid levels, reduced fat accumulation, and mitigated hepatic injury in HLP mice. In addition, PAPS1 demonstrated the capability to increase the levels of three distinct fecal metabolites while simultaneously reducing the levels of eight other fecal metabolites in HLP mice. According to biological detection, PAPS1 reduced the hepatic level of reactive oxygen species (ROS) and pro-inflammatory factors, such as tumor necrosis factor (TNF)-α and interleukin (IL)-1β, -6, -17A, -22, and -23, and increased the expression of anti-inflammatory factor IL-10. Combined with proteomics, Western blot and immunohistochemistry analysis showed that PAPS1 exerted suppressive effects on inflammation and oxidative damage by inhibiting the nuclear factor-κB (NF-κB)/signal transducer and activator of transcription 3 (STAT3) signaling pathway in HLP mice. These findings offer evidence supporting the effectiveness of PAPS1 as a therapeutic agent in reducing lipid levels through its targeting of chronic inflammation.


A Trigeminal Neuropathy From an Inactive Hepatocellular Carcinoma.
Breast, lung, prostate, thyroid, and kidney carcinomas are the primary tumors that are known to have bony metastasis. Hepatocellular carcinoma (HCC) frequently involves the lung and lymph nodes and less commonly the osseous system. Numbness/persistent pain in the distribution of the trigeminal nerve is more likely a neuropathy. The causes are idiopathic(common), unintentional injury to the trigeminal nerve during surgery or trauma, blood vessel pressing the trigeminal nerve, tumor infiltration, multiple sclerosis, and stroke. Unresolved facial pains after conventional treatment should prompt additional investigation to rule out other causes. In this case, we report a trigeminal neuropathy of rare cause, which is a solitary metastasis from an inactive HCC involving the osseous structures.


Establishment and maintenance of human embryonic stem cells on STO, a permanently growing cell line.
Human embryonic stem (hES) cells have been traditionally cultured on primary mouse embryonic fibroblasts (PMEFs). However, though STO cells have some advantages over PMEFs and human embryonic fibroblasts (hEFs) as feeder cells, they have never been used as feeder cells to establish hES cell lines. In this study, three hES cell lines (Miz-hES1, Miz-hES2, and Miz-hES3) were established from inner cell masses (ICM), using STO as feeder cells. The three hES cell lines had normal karyotypes and expressed high levels of alkaline phosphatase (AP), cell surface markers (SSEA-3, SSEA-4, TRA-1-60, and TRA-1-81), and transcription factor Oct-4. After culture on STO cells for 2 yr, hES cells maintained the potential to form derivatives of all three embryonic germ layers. Our results show that STO feeder cells have the potential to support the establishment and maintenance of hES cell lines. In addition, our results suggest that laminin may play an important role in maintaining the undifferentiated proliferation of hES cells.


Skin sensitization data for methyl esters of sulphonic acids-Corrections to published databases required.
Several methyl esters of sulphonic acids are listed in murine local lymph node assay (LLNA) databases, with dose-response data and EC3 values. However, some of these entries are questionable-in one case the chemical tested is not the chemical named in the databases and in others the EC3 value has been derived by extrapolation from data that do not meet the applicability criteria for the approved extrapolation method. To consider how LLNA data came to be attributed to the wrong chemical and to address the inappropriate extrapolated EC3 values. Dose-response data for methyl hexadec-3-enesulphonate (wrongly named as methyl hexadec-1-enesulphonate), two other methyl sulphonates and hexadec-1-ene-1,3-sultone are re-evaluated using the single dose probit extrapolation method (SDPEM). The different reaction chemistry profiles of methyl hexadec-3-enesulphonate and methyl hexadec-1-enesulphonate are discussed. Extrapolated EC3 values for hexadec-1-ene-1,3-sultone are the same by both methods but for the methyl sulphonates the differences are substantial. Current databases should be corrected and further analysed to identify other cases where EC3 values are likely to be unreliable due to inappropriate estimation by extrapolation.


Age-related hearing loss, vitamin B-12, and folate in elderly women.
Hearing impairment is 1 of the 4 most prevalent chronic conditions in the elderly. However, the biological basis of age-related hearing loss is unknown. The objective was to test the hypothesis that age-related hearing loss may be associated with poor vitamin B-12 and folate status. A thorough audiometric assessment was conducted in 55 healthy women aged 60-71 y. Hearing function was determined by the average of pure-tone air conduction thresholds at 0.5, 1, 2, and 4 kHz and was categorized into 2 groups for logistic regression analyses: normal hearing (<20 dB hearing level; n = 44) and impaired hearing (> or = 20 dB hearing level; n = 11). Mean age was the same (65 y) for the normal hearing and impaired hearing groups. Pure-tone averages were inversely correlated with serum vitamin B-12 (r = -0.58, P = 0.0001) and red cell folate (r = -0.37, P = 0.01). Women with impaired hearing had 38% lower serum vitamin B-12 (236 compared with 380 pmol/L, respectively, P = 0.008) and 31% lower red cell folate (425 compared with 619 nmol/L, respectively, P = 0.02) than women with normal hearing. Among participants who did not take supplements containing vitamin B-12 or folate, women with impaired hearing had 48% lower serum vitamin B-12 (156 compared with 302 pmol/L, respectively, P = 0.0007) and 43% lower red cell folate (288 compared with 502 nmol/L, respectively, P = 0.001) than women with normal hearing. Poor vitamin B-12 and folate status may be associated with age-related auditory dysfunction.


Increased coupling of intrinsic networks in remitted depressed youth predicts rumination and cognitive control.
Functional connectivity MRI (fcMRI) studies of individuals currently diagnosed with major depressive disorder (MDD) document hyperconnectivities within the default mode network (DMN) and between the DMN and salience networks (SN) with regions of the cognitive control network (CCN). Studies of individuals in the remitted state are needed to address whether effects derive from trait, and not state or chronic burden features of MDD. fcMRI data from two 3.0 Tesla GE scanners were collected from 30 unmedicated (47% medication naïve) youth (aged 18-23, modal depressive episodes = 1, mean age of onset = 16.2, SD = 2.6) with remitted MDD (rMDD; modal years well = 4) and compared with data from 23 healthy controls (HCs) using four bilateral seeds in the DMN and SN (posterior cingulate cortex (PCC), subgenual anterior cingulate (sgACC), and amygdala), followed by voxel-based comparisons of the whole brain. Compared to HCs, rMDD youth exhibited hyperconnectivities from both PCC and sgACC seeds with lateral, parietal, and frontal regions of the CCN, extending to the dorsal medial wall. A factor analysis reduced extracted data and a PCC factor was inversely correlated with rumination among rMDD youth. Two factors from the sgACC hyperconnectivity clusters were related to performance in cognitive control on a Go/NoGo task, one positively and one inversely. Findings document hyperconnectivities of the DMN and SN with the CCN (BA 8/10), which were related to rumination and sustained attention. Given these cognitive markers are known predictors of response and relapse, hyperconnectivities may increase relapse risk or represent compensatory mechanisms.


Patient outcomes research: seizing the opportunity.
Nurses must take a proactive role in patient outcomes research. Because the patient, not disease, has always been the focus of nursing, nurses are sensitized to the rich but complicated context in which patient outcomes research will be conducted. Nurse researchers are experienced in using complex research models, dealing with ambiguous concepts, grappling with difficult measurement issues, and subscribing to methodological pluralism. These skills are important to success with patient outcomes research. In this article, the opportunities and challenges posed to nurses by patient outcomes research will be addressed. One of the benefits will be a stronger knowledge base for clinical practice and hence patient care.


Laccase from the white-rot fungus Trametes versicolor: cDNA cloning of lcc1 and expression in Pichia pastoris.
A cDNA coding for laccase was isolated from the ligninolytic fungus Trametes versicolor by RNA-PCR. The cDNA corresponds to the gene lcc1, which encodes a laccase isoenzyme of 498 amino-acid residues preceded by a 22-residue signal peptide. The lcc1 cDNA was cloned into the vector pHIL-D2 for expression in Pichia pastoris under the control of the AOX1 promoter. Transformants were found to secrete active recombinant enzyme after induction with methanol. The use of growth medium buffered to pH 6.0 and control of pH during cultivation were found to be important, or even necessary, for obtaining activity in liquid cultures. The effect of exchanging the native secretion signal for the Saccharomyces cerevisiae alpha-factor pre-pro secretion signal was studied by cloning the portion encoding the mature enzyme into the vector pPIC9. The activity obtained for the construct encoding the native laccase signal sequence was found to be seven-fold higher than for the construct encoding the alpha-factor secretion signal. Utilisation of the P. pastoris pep4 mutant strain SMD1168 was found to provide a two-fold higher level of activity compared with P. pastoris GS115.


Differential gene regulation under altered gravity conditions in follicular thyroid cancer cells: relationship between the extracellular matrix and the cytoskeleton.
Extracellular matrix proteins, adhesion molecules, and cytoskeletal proteins form a dynamic network interacting with signalling molecules as an adaptive response to altered gravity. An important issue is the exact differentiation between real microgravity responses of the cells or cellular reactions to hypergravity and/or vibrations. To determine the effects of real microgravity on human cells, we used four DLR parabolic flight campaigns and focused on the effects of short-term microgravity (22 s), hypergravity (1.8 g), and vibrations on ML-1 thyroid cancer cells. No signs of apoptosis or necrosis were detectable. Gene array analysis revealed 2,430 significantly changed transcripts. After 22 s microgravity, the F-actin and cytokeratin cytoskeleton was altered, and ACTB and KRT80 mRNAs were significantly upregulated after the first and thirty-first parabolas. The COL4A5 mRNA was downregulated under microgravity, whereas OPN and FN were significantly upregulated. Hypergravity and vibrations did not change ACTB, KRT-80 or COL4A5 mRNA. MTSS1 and LIMA1 mRNAs were downregulated/slightly upregulated under microgravity, upregulated in hypergravity and unchanged by vibrations. These data indicate that the graviresponse of ML-1 cells occurred very early, within the first few seconds. Downregulated MTSS1 and upregulated LIMA1 may be an adaptive mechanism of human cells for stabilizing the cytoskeleton under microgravity conditions.


Multimodal angiographic assessment of cerebral arteriovenous malformations: a pilot study.
We describe our protocol of three-dimensional (3D) Roadmap intracranial navigation and image fusion for analysis of the angioarchitecture and endovascular treatment of brain arteriovenous malformations (AVMs). We performed superselective catheterization of brain AVMs feeders under 3D-Roadmap navigation. Angiograms of each catheterized artery on two registered orthogonal views were transferred to the imaging workstations, and dedicated postprocessing imaging software allowed automated multiple overlays of the arterial supply of the AVM superselective acquisitions on the global angiogram in angiographic or 3D views and on coregistered MRI datasets. 11 untreated brain AVMs (4 with hemorrhagic presentation) were explored. The superselective acquisitions were performed under 3D-Roadmap navigation in 74 arteries, for a total of 79 targeted arteries. Imaging analysis was available at table side or postoperatively for discussion of the therapeutic strategy. No complications occurred during superselective catheterization. The accuracy of the coregistration of angiogram and MRI was submillimetric after automated mutual information coregistration, with manual re-registration by the physicians. Superselective angiograms acquired under 3D-Roadmap navigation can be postprocessed with multiple overlays. The fluoroscopic navigation under 3D-Roadmapping and the coregistration of 3D rotational angiography, selective angiography, and 3D MR datasets appears reliable with millimeter accuracy, and could be implemented in the critical brain AVM embolization setting to allow refined analysis of AVM angioarchitecture.


Structural characterization and biological evaluation of a clioquinol-ruthenium complex with copper-independent antileukaemic activity.
In this study, we present the synthesis, biological characterization, and first crystal structure of an organometallic-clioquinol complex. Combining ruthenium with the established apoptotic agent and 8-hydroxyquinoline derivative, clioquinol, resulted in a complex that induces caspase-dependent cell death in leukaemia cells. This activity is copper independent and is improved compared to the parent compound, clioquinol. The study of the mode of action reveals that this clioquinol-ruthenium complex does not intercalate between DNA base pairs. Additionally, this clioquinol-ruthenium complex shows proteasome-independent inhibition of the NFκB signalling pathway, with no effects on cell-cycle distribution. These data suggest a mechanism of action that involves a target profile that is different from that for clioquinol alone.


Rhodobacter capsulatus gains a competitive advantage from respiratory nitrate reduction during light-dark transitions.
Rhodobacter capsulatus N22DNAR(+) possesses a periplasmic nitrate reductase and is capable of reducing nitrate to nitrite under anaerobic conditions. In the absence of light this ability cannot support chemoheterotrophic growth in batch cultures. This study investigated the effect of nitrate reduction on the growth of R. capsulatus N22DNAR(+) during multiple light-dark cycles of anaerobic photoheterotrophic/dark chemoheterotrophic growth conditions in carbon-limited continuous cultures. The reduction of nitrate did not affect the photoheterotrophic growth yield of R. capsulatus N22DNAR(+). After a transition from photoheterotrophic to dark chemoheterotrophic growth conditions, the reduction of nitrate slowed the initial washout of a R. capsulatus N22DNAR(+) culture. Towards the end of a period of darkness nitrate-reducing cultures maintained higher viable cell counts than non-nitrate-reducing cultures. During light-dark cycling of a mixed culture, the strain able to reduce nitrate (N22DNAR(+)) outcompeted the strain which was unable to reduce nitrate (N22). The evidence indicates that the periplasmic nitrate reductase activity supports slow growth that retards the washout of a culture during anaerobic chemoheterotrophic conditions, and provides a protonmotive force for cell maintenance during the dark period before reillumination. This translates into a selective advantage during repeated light-dark cycles, such that in mixed culture N22DNAR(+) outcompetes N22. Exposure to light-dark cycles will be a common feature for R. capsulatus in its natural habitats, and this study shows that nitrate respiration may provide a selective advantage under such conditions.


Candida albicans lacking the gene encoding the regulatory subunit of protein kinase A displays a defect in hyphal formation and an altered localization of the catalytic subunit.
The fungal pathogen Candida albicans switches from a yeast-like to a filamentous mode of growth in response to a variety of environmental conditions. We examined the morphogenetic behavior of C. albicans yeast cells lacking the BCY1 gene, which encodes the regulatory subunit of protein kinase A. We cloned the BCY1 gene and generated a bcy1 tpk2 double mutant strain because a homozygous bcy1 mutant in a wild-type genetic background could not be obtained. In the bcy1 tpk2 mutant, protein kinase A activity (due to the presence of the TPK1 gene) was cyclic AMP independent, indicating that the cells harbored an unregulated phosphotransferase activity. This mutant has constitutive protein kinase A activity and displayed a defective germinative phenotype in N-acetylglucosamine and in serum-containing medium. The subcellular localization of a Tpk1-green fluorescent protein (GFP) fusion protein was examined in wild-type, tpk2 null, and bcy1 tpk2 double mutant strains. The fusion protein was observed to be predominantly nuclear in wild-type and tpk2 strains. This was not the case in the bcy1 tpk2 double mutant, where it appeared dispersed throughout the cell. Coimmunoprecipitation of Bcy1p with the Tpk1-GFP fusion protein demonstrated the interaction of these proteins inside the cell. These results suggest that one of the roles of Bcy1p is to tether the protein kinase A catalytic subunit to the nucleus.


Alterations in substrate specificity and physicochemical properties of deoxythymidine kinase of a drug-resistant herpes simplex virus type 1 mutant.
The deoxythymidine kinase (dTK) activity of a 5-methoxymethyldeoxyuridine-resistant mutant (MMdU(r)-20) of herpes simplex virus type 1 was compared with that of the parental wild-type (WT) virus. The dTK activity induced by the mutant was consistently less than that induced by the WT virus, was inhibited by antibody specific for herpes simplex virus dTK, and was more thermostable than the WT dTK. Further, it was inhibited to a lesser degree than the WT dTK by the nucleoside analogs MMdU and arabinosylthymine (araT), which suggests that one of the effects of the mutation was a selective alteration in substrate recognition by the dTK. The loss of ability to inhibit the mutant dTK by E-(2)-5-bromovinyldeoxyuridine was not as great as that seen with araT and MMdU. This agrees well with our previous observation that the MMdU(r)-20 mutant of herpes simplex virus is only partially resistant to this analog, as compared with araT and MMdU (V. Veerisetty and G. A. Gentry, Virology 114:576-579, 1981). [2-(14)C]araT was used to explore further the resistance to araT. Extracts of cells infected with the mutant, although producing a small amount of [(14)C]araTMP, were unable to produce [(14)C]araTTP, in contrast to extracts of cells infected with the WT virus. Both extracts, however, produced [(14)C]dTTP from [(14)C]deoxyribosylthymine. Finally, the ability of the extracts to phosphorylate [(14)C]dTMP was examined. It was found that this activity was greatly reduced relative to dTK activity in the case of the mutant. These findings suggest that a mutation in the dTK polypeptide has affected recognition not only of nucleoside substrates but of the nucleotide substrate dTMP as well, which agrees with the suggestion of Chen et al. that both activities are located on the same polypeptide (M. S. Chen and W. H. Prusoff, J. Biol. Chem. 253:1325-1327, 1978; M. S. Chen, J. Walker, and W. H. Prusoff, J. Biol. Chem. 254:10747-10753, 1979; M. S. Chen, W. P. Summers, J. Walker, W. C. Summers, and W. H. Prusoff, J. Virol. 30:942-945, 1979).


Role of the Bordetella pertussis P.69/pertactin protein and the P.69/pertactin RGD motif in the adherence to and invasion of mammalian cells.
The role of the Bordetella pertussis P.69/pertactin protein in mammalian cell adhesion and invasion was investigated. Salmonella strains expressing surface-associated P.69/pertactin from a chromosomally located prn gene were significantly more invasive than isogenic parental strains. This effect was most pronounced in the poorly invasive, semi-rough S. typhimurium strain LB5010. Escherichia coli K-12 strain HB101 harbouring the plasmid p41869D, which encodes the full-length prn gene under the control of the tac promoter on the broad-host-range plasmid pMMB66EH, was significantly more adhesive to HEp-2 and Chinese Hamster Ovary (CHO) cells growing in culture than E. coli HB101(pMMB66EH). However, the ability of E. coli to invade mammalian cells was not affected by P.69/pertactin expression. P.69/pertactin-mediated adhesiveness of HB101 to HEp-2 and CHO cells was not influenced by the viability of the bacterial cells. However, adherence was markedly reduced when assays were performed for less than 3 h, at 4 degrees C or in the presence of cycloheximide, suggesting the active participation of the eukaryotic cell in bacterial adhesion. Site-directed mutagenesis was used to mutate Asp to Glu in an Arg-Gly-Asp (RGD-->RGE) sequence present in mature P.69/pertactin and the mutated gene was cloned in the same broad-host-range vector (plasmid p41869E). This mutation had no detectable influence on the ability of P.69/pertactin to mediate adhesion of HB101 to HEp-2 or CHO cells. Plasmids p41869D and p41869E were introduced into the bvg-negative B. pertussis strain BP347. Expression of P.69RGD or P.69RGE did not enhance the adhesiveness of BP347 for epithelial (HEp-2 and CHO) cells.


The Simulation-Based Assessment of Pediatric Rapid Response Teams.
To create scenarios of simulated decompensating pediatric patients to train pediatric rapid response teams (RRTs) and to determine whether the scenario scores provide a valid assessment of RRT performance with the hypothesis that RRTs led by intensivists-in-training would be better prepared to manage the scenarios than teams led by nurse practitioners. A set of 10 simulated scenarios was designed for the training and assessment of pediatric RRTs. Pediatric RRTs, comprising a pediatric intensive care unit (PICU) registered nurse and respiratory therapist, led by a PICU intensivist-in-training or a pediatric nurse practitioner, managed 7 simulated acutely decompensating patients. Two raters evaluated the scenario performances and psychometric analyses of the scenarios were performed. The teams readily managed scenarios such as supraventricular tachycardia and opioid overdose but had difficulty with more complicated scenarios such as aortic coarctation or head injury. The management of any particular scenario was reasonably predictive of overall team performance. The teams led by the PICU intensivists-in-training outperformed the teams led by the pediatric nurse practitioners. Simulation provides a method for RRTs to develop decision-making skills in managing decompensating pediatric patients. The multiple scenario assessment provided a moderately reliable team score. The greater scores achieved by PICU intensivist-in-training-led teams provides some evidence to support the validity of the assessment.


Recent developments in antimicrobial and antifouling coatings to reduce or prevent contamination and cross-contamination of food contact surfaces by bacteria.
Illness as the result of ingesting bacterially contaminated foodstuffs represents a significant annual loss of human quality of life and economic impact globally. Significant research investment has recently been made in developing new materials that can be used to construct food contacting tools and surfaces that might minimize the risk of cross-contamination of bacteria from one food item to another. This is done to mitigate the spread of bacterial contamination and resultant foodborne illness. Internet-based literature search tools such as Web of Science, Google Scholar, and Scopus were utilized to investigate publishing trends within the last 10 years related to the development of antimicrobial and antifouling surfaces with potential use in food processing applications. Technologies investigated were categorized into four major groups: antimicrobial agent-releasing coatings, contact-based antimicrobial coatings, superhydrophobic antifouling coatings, and repulsion-based antifouling coatings. The advantages for each group and technical challenges remaining before wide-scale implementation were compared. A diverse array of emerging antimicrobial and antifouling technologies were identified, designed to suit a wide range of food contact applications. Although each poses distinct and promising advantages, significant further research investment will likely be required to reliably produce effective materials economically and safely enough to equip large-scale operations such as farms, food processing facilities, and kitchens.


Cervical cancer screening with clinic-based Pap test versus home HPV test among Somali immigrant women in Minnesota: a pilot randomized controlled trial.
Cervical cancer is more common in the Somali immigrant population than the general population in the United States (US). There are low rates of cervical cancer screening among Somali women. This study compares cervical cancer screening test completion rates for a home human papilloma virus (HPV) test and standard clinic Pap test. Sixty-three Somali immigrant women aged 30-70 years who had not undergone cervical cancer screening within the past 3 years were randomly assigned to a home HPV test group (intervention) or a clinic Pap test group (control). Test completion rates were measured at 3 months. Univariate and multivariate logistic regression models were used to explore factors associated with test completion (intention-to-treat analysis). Participants in the HPV test group were 14 times more likely to complete the test compared to those in the Pap test group (P = 0.0002). Women who reported having friends/family members to talk about cancer screening were approximately three times more likely to complete any screening test than those who did not (P = 0.127) and participants who reported residing in the US longer were more likely to complete a screening test (P = 0.011). Future research should explore the potential of using the home-based HPV test kits as an initial approach to cervical cancer screening. The use of a self-sampling HPV kit has the potential to increase cervical cancer screening in under-served communities in the US.


Characterization of sublethal microcystin-LR exposure in mice.
Microcystin-LR (MCLR) is a potent hepatotoxin produced by the cyanobacterium Microcystis aeruginosa. The histology of acute lethal toxicity has been well characterized, but histology is limited regarding sublethal exposure. Balb/C mice were given a single sublethal dose of MCLR (45 microg/kg) and euthanized at 2, 4, 12, and 24 hours after exposure. Centrilobular to midzonal hepatocellular hypertrophy with loss of cytosolic vacuolation consistent with glycogen depletion occurred at 2 hours. At 4 hours, central lobular hepatocytes exhibited eccentric areas of eosinophilic cytoplasmic condensation that were partially aggregated around the outer nuclear membrane. The areas were weakly positive for cytokeratin and somewhat resembled the Mallory bodies of alcoholic human hepatitis. Small numbers of apoptotic hepatocytes were seen at 24 hours. The toxin was detectable by immunohistochemistry (IHC) as early as 2 hours and was colocalized with the areas of hepatocellular hypertrophy. Intense nuclear staining occurred at 4 hours; this was no longer evident after 12 hours. Strong staining of apoptotic bodies occurred at 24 hours. Mice that received two daily doses had a marked increase in apoptotic hepatocytes in the centrilobular areas. Lesions at four and seven doses consisted of marked hepatocytomegaly and karyomegaly with parenchymal disarray and cytosolic vacuolation. IHC revealed diffuse staining throughout the liver parenchyma consistent with toxin accumulation. An anti-MCLR monoclonal antibody detected bands at the 40-kDa mark in nuclear extracts that were identified as protein phosphatases 1 and 2A by western blotting, consistent with a covalent interaction between MCLR and nuclear protein phosphatases.


[The validity and reliability of the Social Communication Questionnaire- Turkish form in autistics aged 4-18 years].
The Social Communication Questionnaire (SCQ) is a valid and reliable 40- item scale used to assess of pervasive developmental disorders (PDDs). The aim of this study was to determine the validity and reliability of the SCQ-Turkish Form (SCQ-TF). The study included 100 children and adolescents aged 4-18 years 50 were diagnosed as PDD and 50 were diagnosed with intellectual disability (ID) based on DSM-IV-TR criteria. The consistency, test-retest reliability, content validity, and discriminant validity of SCQ-TF for the groups in the study sample were evaluated. SCQ-TF was compared to the Childhood Autism Rating Scale (CARS), Autism Behavioural Checklist (ABC), and Clinical Global Impression Severity of Illness (CGI-SI). The most appropriate SCQ-TF cut-off point was determined via ROC analysis. The 4-factor structure of SCQ-TF accounted for 43% of the observed total variance. Correlations between the Social Communication Questionnaire -Turkish Form and the other measures were significant. The Cronbach's alpha value for the SCQ-TF total score was 0.80. Intraclass correlation coefficient (ICC) varied between 0.87 and 0.96, and the cut-off point was 15. The findings show that SCQ-TF is valid and reliable for use in Turkey in those aged 4-18 years.


Quantitation of urinary porphyrins by use of second-derivative spectroscopy.
We describe a simple, rapid procedure for measuring porphyrins in urine. After acidification of a urine specimen with hydrochloric acid, the second derivative of the absorption spectrum is recorded in the region of the Soret band maximum. The amplitude of the deflection is linearly related to porphyrin concentration, and the point at which the second derivative minimum occurs provides an estimate of the ratio of uroporphyrin to coproporphyrin in the sample. Nonspecific interference is not significant and reproducibility is excellent. Porphyrin excretion was studied in 24-h urine collections from men and non-pregnant as well as pregnant women. Median daily porphyrin excretion (nmol) was: men, 65.0; non-pregnant women, 60.1; and third-trimester pregnant women, 175.0. Distribution of these data was significantly skewed; upper and lower 2.5 percentile limits are given.


Use of a decision aid to improve treatment decisions in osteoporosis: the osteoporosis choice randomized trial.
Poor adherence to therapy, perhaps related to unaddressed patient preferences, limits the effectiveness of osteoporosis treatment in at-risk women. A parallel patient-level randomized trial in primary care practices was performed. Eligible postmenopausal women with bone mineral density T-scores less than -1.0 and not receiving bisphosphonate therapy were included. In addition to usual primary care, intervention patients received a decision aid (a tailored pictographic 10-year fracture risk estimate, absolute risk reduction with bisphosphonates, side effects, and out-of-pocket cost), and control patients received a standard brochure. Knowledge transfer, patient involvement in decision-making, and rates of bisphosphonate start and adherence were studied. Data came from medical records, post-visit written and 6-month phone surveys, video recordings of clinical encounters, and pharmacy prescription profiles. A total of 100 patients (range of 10-year fracture risk, 6%-60%) were allocated randomly to receive the decision aid (n=52) or usual care (n=48). Patients receiving the decision aid were 1.8 times more likely to correctly identify their 10-year fracture risk (49% vs 28%; 95% confidence interval [CI], 1.03-3.2) and 2.7 times more likely to identify their estimated risk reduction with bisphosphonates (43% vs 16%; 95% CI, 1.3-5.7). Patient involvement improved with the decision aid by 23% (95% CI, 13.6-31.4). Bisphosphonates were started by 44% of patients receiving the decision aid and 40% of patients receiving usual care. Adherence at 6 months was similarly high across both groups, but the proportion with more than 80% adherence was higher with the decision aid (n=23 [100%] vs n=14 [74%]; P = .009). A decision aid improved the quality of clinical decisions about bisphosphonate therapy in at-risk postmenopausal women, did not affect start rates, and may have improved adherence.


Near-continuous, noninvasive blood pressure monitoring in the out-of-hospital setting.
This study was conducted to test out-of-hospital performance of a noninvasive radial artery tonometry device to assess blood pressure (BP), providing readings every 10-12 seconds. The primary objective was to determine the correlation between noninvasive BPs calculated with radial artery tonometry and standard oscillometric cuff methods. The secondary objective was to determine whether the difference observed between the two techniques was consistent over the range of BPs measured. This prospective trial enrolled adults transported by helicopter (n = 9 patients), fixed-wing airplane (n = 1), or ground vehicle (n = 10) of a single transport service. Patients had BP assessed simultaneously, by both standard automatic cuff and radial artery tonometry device, every 5 minutes. Data were assessed with correlation coefficients, and Bland-Altman techniques were utilized to assess for bias over the range of mean arterial pressures (MAPs) encountered. For all tests, p was set at 0.05. No major problem with radial artery tonometry device field performance was noted. There were 139 pairs of MAP assessments in 20 patients. The correlation coefficient for the two assessment modalities was 0.96. Bland-Altman bias plot and Pitman's test (p = 0.11) revealed good correlation between the two assessment mechanisms over the entire range of MAPs (42 to 163 mm Hg) encountered in the study. The radial artery tonometry device provided MAP assessments that were highly correlated with readings from a standard oscillometric device. The radial artery tonometry device performed well in a variety of patient types and in multiple transport vehicles, and there was no sign that its performance was adversely affected by the out-of-hospital setting.


The roles of reward, default, and executive control networks in set-shifting impairments in schizophrenia.
Patients with schizophrenia (SZ) show deficits on tasks of rapid reinforcement learning, like probabilistic reversal learning (PRL), but the neural bases for those impairments are not known. Recent evidence of relatively intact sensitivity to negative outcomes in the ventral striatum (VS) in many SZ patients suggests that PRL deficits may be largely attributable to processes downstream from feedback processing, involving both the activation of executive control task regions and deactivation of default mode network (DMN) components. We analyzed data from 29 chronic SZ patients and 21 matched normal controls (NCs) performing a PRL task in an MRI scanner. Subjects were presented with eight pairs of fractal stimuli, for 50 trials each. For each pair, subjects learned to choose the more frequently-rewarded (better) stimulus. Each time a criterion was reached, the better stimulus became the worse one, and the worse became the better. Responses to feedback events were assessed through whole-brain and regions-of-interest (ROI) analyses in DMN. We also assessed correlations between BOLD signal contrasts and clinical measures in SZs. Relative to NCs, SZ patients showed comparable deactivation of VS in response to negative feedback, but reduced deactivation of DMN components including medial prefrontal cortex (mPFC). The magnitudes of patients' punishment-evoked deactivations in VS and ventromedial PFC correlated significantly with clinical ratings for avolition/anhedonia. These findings suggest that schizophrenia is associated with a reduced ability to deactivate components of default mode networks, following the presentation of informative feedback and that motivational deficits in SZ relate closely to feedback-evoked activity in reward circuit components. These results also confirm a role for ventrolateral and dorsomedial PFC in the execution of response-set shifts.


Skin-sparing mastectomy with immediate breast and nipple reconstruction: a new technique of nipple reconstruction.
Background. Most women with breast cancer today can be managed with breast conservation; however, some women still require mastectomy for treatment of their disease. Skin-sparing mastectomy (SSM) with immediate reconstruction has emerged as a favorable option for many of these patients. The authors combined the SSM technique with the preservation of a small part of the areola with immediate nipple together with with breast reconstruction. Methods. In an 8-year-period 155 female patients (age: 20-52 years old; mean age: 37.5 years) with extensive ductal intraepithelial neoplasia (DIN) or invasive breast cancer were treated with areola skin sparing mastectomy with immediate nipple and breast reconstruction. Patients were followed up prospectively by the breast surgeon, the plastic surgeon, and the oncologist for complications and recurrence. Results. After treatment, only 2 cases (1.29%) had a local recurrence. 8 out of 155 (5.5%) patients developed early complications (infections, seroma, haematoma), and 5 out of 155 patients (3.22%) developed delayed complications (implant rotation, aestethic deterioration) in the post operative time period. The final aesthetic outcome was judged as positive in 150 out of 155 patients (96.78%). Conclusion. In our experience, immediate nipple reconstruction after skin-sparing mastectomy is a technically feasible procedure which can give excellent cosmetic results.


Identification of a new motif required for the 3'-5' exonuclease activity of Escherichia coli DNA polymerase I (Klenow fragment): the RRRY motif is necessary for the binding of single-stranded DNA substrate and the template strand of the mismatched duplex.
The Klenow fragment of Escherichia coli DNA polymerase I houses catalytic centers for both polymerase and 3'-5' exonuclease activities that are separated by about 35 A. Upon the incorporation of a mismatched nucleotide, the primer terminus is transferred from the polymerase site to an exonuclease site designed for excision of the mismatched nucleotides. The structural comparison of the binary complexes of DNA polymerases in the polymerase and the exonuclease modes, together with a molecular modeling of the template strand overhang in Klenow fragment, indicated its binding in the region spanning residues 821-824. Since these residues are conserved in the "A" family DNA polymerases, we have designated this region as the RRRY motif. The alanine substitution of individual amino acid residues of this motif did not change the polymerase activity; however, the 3'-5' exonuclease activity was reduced 2-29-fold, depending upon the site of mutation. The R821A and R822A/Y824A mutant enzymes showed maximum cleavage defect with single-stranded DNA, mainly due to a large decrease in the ssDNA binding affinity of these enzymes. Mismatch removal by these enzymes was only moderately affected. However, data from the exonuclease-polymerase balance assays with mismatched template-primer suggest that the mutant enzymes are defective in switching mismatched primer from the polymerase to the exonuclease site. Thus, the RRRY motif provides a binding track for substrate ssDNA and for nonsubstrate single-stranded template overhang, in a polarity-dependent manner. This binding then facilitates cleavage of the substrate at the exonuclease site.


Relation between plasma ascorbic acid and mortality in men and women in EPIC-Norfolk prospective study: a prospective population study. European Prospective Investigation into Cancer and Nutrition.
Ascorbic acid (vitamin C) might be protective for several chronic diseases. However, findings from prospective studies that relate ascorbic acid to cardiovascular disease or cancer are not consistent. We aimed to assess the relation between plasma ascorbic acid and subsequent mortality due to all causes, cardiovascular disease, ischaemic heart disease, and cancer. We prospectively examined for 4 years the relation between plasma ascorbic acid concentrations and mortality due to all causes, and to cardiovascular disease, ischaemic heart disease, and cancer in 19 496 men and women aged 45-79 years. We recruited individuals by post using age-sex registers of general practices. Participants completed a health and lifestyle questionnaire and were examined at a clinic visit. They were followed-up for causes of death for about 4 years. Individuals were divided into sex-specific quintiles of plasma ascorbic acid. We used the Cox proportional hazard model to determine the effect of ascorbic acid and other risk factors on mortality. Plasma ascorbic acid concentration was inversely related to mortality from all-causes, and from cardiovascular disease, and ischaemic heart disease in men and women. Risk of mortality in the top ascorbic acid quintile was about half the risk in the lowest quintile (p<0.0001). The relation with mortality was continuous through the whole distribution of ascorbic acid concentrations. 20 micromol/L rise in plasma ascorbic acid concentration, equivalent to about 50 g per day increase in fruit and vegetable intake, was associated with about a 20% reduction in risk of all-cause mortality (p<0.0001), independent of age, systolic blood pressure, blood cholesterol, cigarette smoking habit, diabetes, and supplement use. Ascorbic acid was inversely related to cancer mortality in men but not women. Small increases in fruit and vegetable intake of about one serving daily has encouraging prospects for possible prevention of disease.


A nonheme high-spin ferrous pool in mitochondria isolated from fermenting Saccharomyces cerevisiae.
Mössbauer spectroscopy was used to detect pools of Fe in mitochondria from fermenting yeast cells, including those consisting of nonheme high-spin (HS) Fe(II) species, Fe(III) nanoparticles, and mononuclear HS Fe(III) species. At issue was whether these species were located within mitochondria or on their exterior. None could be removed by washing mitochondria extensively with ethylene glycol tetraacetic acid or bathophenanthroline sulfonate (BPS), Fe(II) chelators that do not appear to penetrate mitochondrial membranes. However, when mitochondrial samples were sonicated, BPS coordinated the Fe(II) species, forming a low-spin Fe(II) complex. This treatment also diminished the levels of both Fe(III) species, suggesting that all of these Fe species are encapsulated by mitochondrial membranes and are protected from chelation until membranes are disrupted. 1,10-Phenanthroline is chemically similar to BPS but is membrane soluble; it coordinated nonheme HS Fe(II) in unsonicated mitochondria. Further, the HS Fe(III) species and nanoparticles were not reduced by dithionite until the detergent deoxycholate was added to disrupt membranes. There was no correlation between the percentage of nonheme HS Fe(II) species in mitochondrial samples and the level of contaminating proteins. These results collectively indicate that the observed Fe species are contained within mitochondria. Mossbauer spectra of whole cells were dominated by HS Fe(III) features; the remainder displayed spectral features typical of isolated mitochondria, suggesting that the Fe in fermenting yeast cells can be coarsely divided into two categories: mitochondrial Fe and (mostly) HS Fe(III) ions in one or more non-mitochondrial locations.


Evaluation of Prognostic Factors and Proposed Changes to the Modified Tokuhashi Score in Patients With Spinal Metastases From Breast Cancer.
A retrospective study of all patients with histologically confirmed breast cancer spinal metastases presenting to a single institution between May 2001 and April 2012. The aim of this study was to investigate whether the 2014mT is more accurate than the 2005mT. The commonly used 2005 modified Tokuhashi score (2005mT) has become more inaccurate as oncologists move toward treating tumors according to their molecular and genomic profile, rather than their tissue-of-origin. In attempts to improve the accuracy of the 2005mT, a revised score (2014mT) was published, suggesting that hormone receptor negative and triple-negative breast cancer patients be given a modified Tokuhashi histological score of 3 rather than 5. Demographic characteristics, tumor receptor status, clinical findings in relation to the primary tumor and its metastases, and actual survival time were collated. The 2005mT was compared with the 2014mT. Univariate and multivariate Cox regression analyses were used to evaluate the influence of each parameter on survival, and receiver operating characteristic curves were used to determine predictive values of each score version. Of the 185 patients included, 32 underwent operative treatment, while 153 were managed nonoperatively for their spinal metastases. The overall cohort had a median survival time of 24 months following the diagnosis of spinal metastases, with a 6-month survival rate of 90%. Hormone, HER2 and triple-negative receptor statuses were significant predictors of poorer survival upon multivariate analysis (P = 0.004, P = 0.007, P < 0.001, and P < 0.001, respectively). Age, the original Tokuhashi score components, previous breast surgery for cancer, previous radiotherapy to the breast, previous radiotherapy to the spine, previous chemotherapy, and previous immunotherapy were not significant. At 6 months, the 2005mT AUROC was 0.62, while that of the 2014mT was 0.64 (P = 0.5394). Tumor histological subtype is crucial when prognosticating the survival of patients with breast cancer spinal metastases. Although the 2014mT was marginally more accurate than the 2005mT, its predictive ability remains poor. 3.


[Some characteristics of the kinetics of enzymatic reactions based on the example of catalase].
A method of kinetic analysis for quickly acting enzymes, which are characterized with substrate inhibition, on the catalase model is proposed. Catalase kinetics was shown to be full described, considering changes in the maximal reaction rate and Michaelis constant, by four parameters instead of two usual ones (Vmax, Km equals const.). The method described makes possible to calculate the change the Michaelis constant in time and to estimate real dependencies of the reaction rate on time and on the substrate concentration. Moreover, the enzyme concentration and its inactivation rate at any reaction moment can be calculated under saturation conditions. It is supposed that experimental dependencies of Km on t and of Vmax on t are the results of residual conformation changes accumulated by the enzyme in the reaction process.


CETA and pharmaceuticals: impact of the trade agreement between Europe and Canada on the costs of prescription drugs.
On a per capita basis, Canadian drug costs are already the second highest in the world after the United States and are among the fastest rising in the Organization for Economic Co-Operation and Development. The Comprehensive Economic and Trade Agreement (CETA) between the European Union (EU) and Canada will further exacerbate the rise in costs by: Committing Canada to creating a new system of patent term restoration thereby delaying entry of generic medicines by up to two years; Locking in Canada's current term of data protection, and creating barriers for future governments wanting to reverse it; Implementing a new right of appeal under the patent linkage system that will create further delays for the entry of generics.CETA will only affect intellectual property rights in Canada-not the EU. This analysis estimates that CETA's provisions will increase Canadian drug costs by between 6.2% and 12.9% starting in 2023. The Canadian government committed to compensating provinces for the rise in costs for their public drug plans. Importantly, this means that people paying out-of-pocket for their drugs or receiving them through private insurance, will be charged twice: once through higher drug costs and once more through their federal taxes.As drug costs continue to grow, there are limited options available for provincial/territorial governments: restrict the choice of medicines in public drug plans; transfer costs to patients who typically are either elderly or sick; or take money from other places in the health system, and threaten the viability of Canada's single payer system. CETA will therefore negatively impact the ability of Canada to offer quality health care.


Hyperthermia-induced enhancement of melphalan activity against a melphalan-resistant human rhabdomyosarcoma xenograft.
The effects of regional hyperthermia (42 degrees C for 70 min) on the antitumor activity of melphalan were examined in athymic mice bearing melphalan-resistant human rhabdomyosarcoma (TE-671 MR) xenografts growing in the right hind limb, and results were compared with similar studies of melphalan-sensitive (TE-671) parent xenografts. Melphalan alone at a dose of 36 mg/m2 (0.5 of the 10% lethal dose) produced growth delays of 4.1 to 10.2 days in TE-671 MR xenografts and 21.8 to 28.7 days in TE-671, respectively. Hyperthermia alone produced growth delays of 0.9 days in TE-671 MR xenografts and 0.8 days in TE-671. Combination therapy with melphalan and hyperthermia produced growth delays of 7.2 to 13.3 days in TE-671 MR xenografts and 34.3 to 42.8 days in TE-671, respectively, representing a mean thermal enhancement ratio of 1.7 in TE-671 MR and 1.5 in TE-671. Measurement of glutathione levels in TE-671 MR xenografts following treatment with melphalan, hyperthermia, or melphalan plus hyperthermia revealed significant reductions in glutathione content with the nadir (60% of control values) seen 6 h following treatment. Glutathione levels in TE-671 xenografts following identical therapy revealed no differences from control values. Hyperthermia plus melphalan did not result in a higher tumor-to-plasma melphalan ratio compared with treatment with melphalan alone in either TE-671 MR or TE-671 xenografts. These studies suggest that heat-induced alterations in tumor glutathione or melphalan levels are not responsible for the increase in melphalan activity produced by hyperthermia. Combination therapy with melphalan plus regional hyperthermia offers promise for treatment of melphalan-resistant neoplasms.


Resting-State Functional MRI and PET Imaging as Noninvasive Tools to Study (Ab)Normal Neurodevelopment in Humans and Rodents.
Neurodevelopmental disorders (NDDs) are a group of complex neurologic and psychiatric disorders. Functional and molecular imaging techniques, such as resting-state functional magnetic resonance imaging (rs-fMRI) and positron emission tomography (PET), can be used to measure network activity noninvasively and longitudinally during maturation in both humans and rodent models. Here, we review the current knowledge on rs-fMRI and PET biomarkers in the study of normal and abnormal neurodevelopment, including intellectual disability (ID; with/without epilepsy), autism spectrum disorder (ASD), and attention deficit hyperactivity disorder (ADHD), in humans and rodent models from birth until adulthood, and evaluate the cross-species translational value of the imaging biomarkers. To date, only a few isolated studies have used rs-fMRI or PET to study (abnormal) neurodevelopment in rodents during infancy, the critical period of neurodevelopment. Further work to explore the feasibility of performing functional imaging studies in infant rodent models is essential, as rs-fMRI and PET imaging in transgenic rodent models of NDDs are powerful techniques for studying disease pathogenesis, developing noninvasive preclinical imaging biomarkers of neurodevelopmental dysfunction, and evaluating treatment-response in disease-specific models.


Self reported symptoms and inhibition of acetylcholinesterase activity among Kenyan agricultural workers.
This study was part of the East African pesticides project. The general objective was to assess health hazards posed by handling, storage, and use of pesticides, on agricultural estates and small farms with a view to developing strategies for prevention and control of pesticide poisoning. The aim of this paper is to describe the prevalence of symptoms in this population, to relate levels of inhibition to reported symptoms and evaluate at which levels of inhibition symptoms become increased. Complete data were available for 256 exposed subjects and 152 controls from four regions in Kenya. A structured questionnaire on symptoms experienced at the time of interview was given to all subjects and controls. Information was also obtained on sex, age, main occupation, and level of education. Symptoms reported during the high exposure period, were initially clustered in broader symptom categories from reference literature on health effects of pesticides that inhibit cholinesterase (organophosphate and carbamate). Prevalence ratios were estimated for symptoms with changes in cholinesterase activity in serum. Symptom prevalence in exposed subjects was higher during the high exposure period than the low exposure period, although these differences were not significant. Interestingly, a clear and significant change in symptoms prevalence was found in the controls with a higher prevalence in the low exposure period. Analysis of the relation between cholinesterase inhibition and symptoms showed that prevalence ratios were significantly > 1 for respiratory, eye, and central nervous system symptoms for workers with > 30% inhibition. Similar results were found for analyses with the actual level of acetylcholinesterase activity. The results suggest the presence of a relation between exposure and acetylcholinesterase inhibition, acetylcholinesterase activity, and respiratory, eye, and central nervous system symptoms. Increased symptom prevalence was found at acetylcholinesterase activities generally considered to be non-adverse.


Exploration of the role of the subodontoblastic layer in odontoblast-like cell differentiation after tooth drilling using Nestin-enhanced green fluorescent protein transgenic mice.
Original odontoblasts and regenerated odontoblast-like cells (OBLCs) may differently regulate Nestin expression. This study aimed to investigate the role of the subodontoblastic layer (SOBL) using green fluorescent protein (GFP) reactivity in the process of OBLC differentiation after tooth drilling in Nestin-enhanced GFP transgenic mice. A groove-shaped cavity was prepared on the mesial surface of the maxillary first molars of 5- or 6-week-old mice under deep anesthesia. Immunohistochemical staining for Nestin and GFP and Nestin in situ hybridization were conducted on the sections obtained at 1-14 days postoperative. Odontoblasts showed intense endogenous Nestin protein and mRNA expression, whereas the coronal SOBL cells showed a Nestin-GFP-positive reaction in the control groups. The injured odontoblasts had significantly decreased Nestin immunoreactivity as well as decreased expression of Nestin mRNA 1-2 days after the injury; subsequently, newly differentiated OBLCs were arranged along the pulp-dentin border, with significantly increased Nestin expression as well as increased expression of Nestin mRNA on days 3-5 to form reparative dentin. Nestin-GFP-positive cells at the pulp-dentin border significantly increased in number on days 1 and 2. GFP(+)/Nestin(+) and GFP(-)/Nestin(+) cells were intermingled in the newly differentiated OBLCs. The commitment of Nestin-GFP-positive cells into Nestin-positive OBLCs suggests that the restriction of endogenous Nestin protein and mRNA expression in the static SOBL cells was removed by exogenous stimuli, resulting in their migration along the pulp-dentin border and their differentiation into OBLCs.


Therapeutic benefits of infrared (810-nm) diode laser macular grid photocoagulation in prophylactic treatment of nonexudative age-related macular degeneration: two-year results of a randomized pilot study.
This pilot study collected preliminary information on the effectiveness and safety of infrared (810-nm) diode laser macular grid photocoagulation in patients with nonexudative age-related macular degeneration (AMD). Results from this pilot study were used in designing a larger, multicenter, randomized clinical trial. A multicenter, randomized, controlled, clinical trial. A total of 229 eyes of 152 patients with AMD were enrolled in the pilot study. Seventy-five patients with 1 eye eligible (75 eyes) were enrolled in the unilateral arm of the study; 77 patients with both eyes eligible (154 eyes) were enrolled in the bilateral arm of the study. In the unilateral study arm, 32 eyes were randomized to the observation group, 27 eyes were treated with visible endpoint burns, and 16 eyes were treated with invisible endpoint (subthreshold) lesions. In the bilateral study arm, 77 eyes were in the observation group, 36 eyes were treated with visible burns, and 41 eyes were treated with subthreshold (invisible) lesions. Eyes were treated with infrared (810-nm) diode laser macular grid photocoagulation using either visible burns or subthreshold (invisible) lesions and compared to eyes receiving no treatment. Reduction of drusen, change in visual acuity, and rate of choroidal neovascularization (CNV) membrane formation. At 12 months after treatment, 62% of eyes treated with visible burns had a clinically significant reduction in drusen, whereas this proportion (65%) was reached in 18 months for eyes treated with subthreshold lesions. At 24 months' follow-up, treated eyes had a significant reduction in drusen compared to observation eyes (P < 0.0001). Visual acuity was significantly improved in treated eyes at 12, 18, and 24 months compared to observation eyes (P < 0.001). Choroidal neovascularization formation was similar in treated and observation eyes through 24 months' follow-up. Complications included CNV associated with six eyes treated with visible burns and a juxtafoveal laser scar in one eye treated with visible burns. Infrared (810-nm) diode laser macular grid photocoagulation in patients with nonexudative AMD significantly reduces drusen levels (P < 0.0001) and significantly improves visual acuity (P < 0.001) when either visible endpoint burns or subthreshold endpoint lesions are used. Complications were fewer using subthreshold endpoint lesions. A larger, multicenter, prospective clinical trial with longer follow-up is needed to determine the efficacy of treatment in reducing the rate of CNV formation. Data from this clinical pilot study have been used to design the Prophylactic Treatment of AMD Trial (PTAMD), a multicenter, randomized, prospective clinical trial currently in progress comparing subthreshold (invisible) treatment to observation in eyes with nonexudative AMD.