Neurophysiological monitoring in the endovascular therapy of aneurysms.
Endovascular aneurysm therapy has associated risks of ischemic complications. We undertook this study to evaluate the efficacy of neurophysiological monitoring (NPM) techniques in the detection of ischemic changes that may be seen during endovascular treatment of cerebral aneurysms. Thirty-five patients underwent NPM during endovascular treatment of cerebral aneurysms. The patients underwent a total of 50 endovascular procedures, including balloon test occlusion (19 patients), GDC embolization (22 patients), and permanent vessel occlusion (nine patients). NPM included electroencephalography, somatosensory evoked potentials, and/or brain stem auditory evoked potentials, depending on the location of the aneurysm. NPM changes were seen in nine (26%) of 35 patients and altered the management in five (14%) of 35 patients. In three of the five cases, NPM changes were observed without corresponding neurologic physical examination changes after balloon test occlusion (performed while the patients were under general anesthesia in two cases). In the two other cases in which NPM changes altered management, ischemia was detected at the time of intra-aneurysmal therapy while the patients were under general anesthesia. Overall, 18 of 35 patients underwent a total of 19 balloon test occlusion procedures. Of the 17 remaining patients, 13 underwent aneurysm coiling, two were not treated because of inability to safely place coils, and two were treated for distal aneurysms. Two patients developed transient neurologic deficits without concurrent NPM changes, representing false-negative NPM test results. NPM is a valuable adjunct to endovascular treatment of cerebral aneurysms. Our study suggests that these monitoring techniques may reduce ischemic complications and can be used to help guide therapeutic decisions.


A prospective study of plasma total cysteine and risk of breast cancer.
Cysteine is the precursor of glutathione, a powerful intracellular antioxidant and an important detoxifying agent of carcinogens. However, data relating plasma total cysteine to breast cancer risk are sparse. We conducted a prospective nested case-control study among 32826 women in the Nurses' Health Study who provided blood specimens during 1989-1990. Through 1996, a total of 712 incident breast cancer cases were identified and individually matched to 712 controls by year of birth, time of day that blood was drawn, fasting status, month of blood sampling, recent use of postmenopausal hormones at the time of blood collection, and menopausal status. Conditional logistic regression with adjustment for other risk factors for breast cancer was used to estimate the relative risks and 95% confidence intervals of breast cancer by levels of plasma total cysteine. Higher plasma total cysteine concentrations were significantly associated with a lower risk of breast cancer. For women in the highest quintile of plasma total cysteine, compared with those in the lowest quintile, the multivariate relative risk was 0.44 (95% confidence interval, 0.26-0.74). This association was dose dependent (P for trend = 0.002) and independent of plasma measures of folate, vitamin B(6), vitamin B(12), and total homocysteine. The inverse association between plasma total cysteine concentrations and risk of breast cancer was not significantly modified by other risk factors for breast cancer, except that a stronger association was observed among women who were leaner. The findings from this prospective study suggest that higher plasma concentrations of total cysteine predict a reduced risk of breast cancer. Cysteine or its precursors might have the potential to be chemopreventive against breast cancer.


Posttraumatic stress disorder symptoms in resident physicians related to their internship.
To evaluate whether stresses experienced by physicians during internship are associated with posttraumatic stress disorder (PTSD) symptoms, the authors administered a standardized questionnaire to assess PTSD symptoms, depressive and anxiety symptoms, and social supports to 212 residents. Thirteen percent of residents met diagnostic criteria for PTSD, including 20% of the women and 9% of the men. PTSD symptoms were significantly more frequent in residents who were single or divorced at the time the questionnaire was administered. There were significant positive correlations between PTSD symptoms and symptoms of both current anxiety and depression, and a significant negative correlation existed between social support and PTSD symptoms. PTSD symptoms attributed to stresses associated with internship, which may be deleterious to physician well-being and potentially to patient care, and suggest the importance of making major changes in postgraduate medical training.


[Current situation of the most frequent zoonosis in the world].
Zoonoses are at the present time more important than ever due to their magnitude and impact. The international trade in animals, products and sub products, as well as the intense travel of people around the world, represent risks of dissemination of infectious diseases, and are the reason for a new age of emerging and reemerging zoonotic diseases. Under these conditions, public health and animal health authorities are obliged to work together in order to get more efficient control programs. In this paper the actual situation of some important emerging and reemerging zoonoses is analyzed, including: anthrax, rabies, tuberculosis, brucellosis, cysticercosis, echinococcosis, hanta virus, Hendra and Nipah virus. Particular attention is given to leptospirosis, due to the fact that it is considered by WHO and OIE as the widest spread zoonotic disease in the world. Zoonoses caused by ingestion of animal food products are discussed. They are responsible for the death of almost 2.2 million people. Bacteria of genus salmonella and campylobacter are considered. Some recommendations are given for the control and prevention of zoonoses, emphasizing the "One Health" concept.


Simulated Microgravity Influences VEGF, MAPK, and PAM Signaling in Prostate Cancer Cells.
Prostate cancer is one of the leading causes of cancer mortality in men worldwide. An unusual but unique environment for studying tumor cell processes is provided by microgravity, either in space or simulated by ground-based devices like a random positioning machine (RPM). In this study, prostate adenocarcinoma-derived PC-3 cells were cultivated on an RPM for time periods of 3 and 5 days. We investigated the genes associated with the cytoskeleton, focal adhesions, extracellular matrix, growth, survival, angiogenesis, and metastasis. The gene expression of signaling factors of the vascular endothelial growth factor (VEGF), mitogen-activated protein kinase (MAPK), and PI3K/AKT/mTOR (PAM) pathways was investigated using qPCR. We performed immunofluorescence to study the cytoskeleton, histological staining to examine the morphology, and a time-resolved immunofluorometric assay to analyze the cell culture supernatants. When PC-3 cells were exposed to simulated microgravity (s-µg), some cells remained growing as adherent cells (AD), while most cells detached from the cell culture flask bottom and formed multicellular spheroids (MCS). After 3-day RPM exposure, PC-3 cells revealed significant downregulation of the VEGF, SRC1, AKT, MTOR, and COL1A1 gene expression in MCS, whereas FLT1, RAF1, MEK1, ERK1, FAK1, RICTOR, ACTB, TUBB, and TLN1 mRNAs were not significantly changed. ERK2 and TLN1 were elevated in AD, and FLK1, LAMA3, COL4A5, FN1, VCL, CDH1, and NGAL mRNAs were significantly upregulated in AD and MCS after 3 days. After a 5-day culture in s-µg, the PC-3 cells showed significant downregulations of VEGF mRNA in AD and MCS, and FN1, CDH1, and LAMA3 in AD and SCR1 in MCS. In addition, we measured significant upregulations in FLT1, AKT, ERK1, ERK2, LCN2, COL1A1, TUBB, and VCL mRNAs in AD and MCS, and increases in FLK1, FN1, and COL4A5 in MCS as well as LAMB2, CDH1, RAF1, MEK1, SRC1, and MTOR mRNAs in AD. FAK1 and RICTOR were not altered by s-µg. In parallel, the secretion rate of VEGFA and NGAL proteins decreased. Cytoskeletal alterations (F-actin) were visible, as well as a deposition of collagen in the MCS. In conclusion, RPM-exposure of PC-3 cells induced changes in their morphology, cytoskeleton, and extracellular matrix protein synthesis, as well as in their focal adhesion complex and growth behavior. The significant upregulation of genes belonging to the PAM pathway indicated their involvement in the cellular changes occurring in microgravity.


Evaluations for in vitro correlates of immunogenicity of inactivated influenza a H5, H7 and H9 vaccines in humans.
Serum antibody responses in humans to inactivated influenza A (H5N1), (H9N2) and A (H7) vaccines have been varied but frequently low, particularly for subunit vaccines without adjuvant despite hemagglutinin (HA) concentrations expected to induce good responses. To help understand the low responses to subunit vaccines, we evaluated influenza A (H5N1), (H9N2), (H7N7) vaccines and 2009 pandemic (H1N1) vaccines for antigen uptake, processing and presentation by dendritic cells to T cells, conformation of vaccine HA in antibody binding assays and gel analyses, HA titers with different red blood cells, and vaccine morphology in electron micrographs (EM). Antigen uptake, processing and presentation of H5, H7, H9 and H1 vaccine preparations evaluated in humans appeared normal. No differences were detected in antibody interactions with vaccine and matched virus; although H7 trimer was not detected in western blots, no abnormalities in the conformation of the HA antigens were identified. The lowest HA titers for the vaccines were <1:4 for the H7 vaccine and 1:661 for an H9 vaccine; these vaccines induced the fewest antibody responses. A (H1N1) vaccines were the most immunogenic in humans; intact virus and virus pieces were prominent in EM. A good immunogenic A (H9N2) vaccine contained primarily particles of viral membrane with external HA and NA. A (H5N1) vaccines intermediate in immunogenicity were mostly indistinct structural units with stellates; the least immunogenic A (H7N7) vaccine contained mostly small 5 to 20 nm structures. Antigen uptake, processing and presentation to human T cells and conformation of the HA appeared normal for each inactivated influenza A vaccine. Low HA titer was associated with low immunogenicity and presence of particles or split virus pieces was associated with higher immunogenicity.


Increased liver AGEs induce hepatic injury mediated through an OST48 pathway.
The protein oligosaccharyltransferase-48 (OST48) is integral to protein N-glycosylation in the endoplasmic reticulum (ER) but is also postulated to act as a membrane localised clearance receptor for advanced glycation end-products (AGE). Hepatic ER stress and AGE accumulation are each implicated in liver injury. Hence the objective of this study was to increase the expression of OST48 and examine the effects on hepatic function and structure. Groups of 8 week old male mice (n = 10-12/group) over-expressing the gene for OST48, dolichyl-diphosphooligosaccharide-protein glycosyltransferase (DDOST+/-), were followed for 24 weeks, while randomised to diets either low or high in AGE content. By week 24 of the study, either increasing OST48 expression or consumption of high AGE diet impaired liver function and modestly increased hepatic fibrosis, but their combination significantly exacerbated liver injury in the absence of steatosis. DDOST+/- mice had increased both portal delivery and accumulation of hepatic AGEs leading to central adiposity, insulin secretory defects, shifted fuel usage to fatty and ketoacids, as well as hepatic glycogen accumulation causing hepatomegaly along with hepatic ER and oxidative stress. This study revealed a novel role of the OST48 and AGE axis in hepatic injury through ER stress, changes in fuel utilisation and glucose intolerance.


Current concepts in the diagnosis and treatment of brain tumours in dogs and cats.
Intracranial tumours occur relatively frequently in dogs, and less commonly in cats. With the availability of computed tomography (CT) and magnetic resonance imaging, more accurate determination of the location and extent of brain tumours in companion animals has become possible. Following these advances in imaging, precise CT-guided stereotactic techniques for both tumour biopsy and intratumoral drug delivery have been developed for use in cats and dogs. Also, tumour identification methods, such as crush preparation examination, have facilitated rapid tumour identification. The use of improved diagnostic techniques has resulted in an increasing demand for effective therapies for brain tumours. While surgical removal and irradiation remain important treatment considerations in the management of brain tumours of cats and dogs, the development of gene therapy strategies for treatment of intracranial tumours offers much promise, although research in this area is still at an early stage.


Apolipoprotein A-I concentrations and risk of coronary artery disease: A Mendelian randomization study.
Apolipoprotein A-I (apoA-I) infusions represent a potential novel therapeutic approach for the prevention of coronary artery disease (CAD). Although circulating apoA-I concentrations inversely associate with risk of CAD, the evidence base of this representing a causal relationship is lacking. The aim was to assess the causal role of apoA-I using human genetics. We identified a variant (rs12225230) in APOA1 locus that associated with circulating apoA-I concentrations (p < 5 × 10-8) in 20,370 Finnish participants, and meta-analyzed our data with a previous GWAS of apoA-I. We obtained genetic estimates of CAD from UK Biobank and CARDIoGRAMplusC4D (totaling 122,733 CAD cases) and conducted a two-sample Mendelian randomization analysis. We compared our genetic findings to observational associations of apoA-I with risk of CAD in 918 incident CAD cases among 11,535 individuals from population-based prospective cohorts. ApoA-I was associated with a lower risk of CAD in observational analyses (HR 0.81; 95%CI: 0.75, 0.88; per 1-SD higher apoA-I), with the association showing a dose-response relationship. Rs12225230 associated with apoA-I concentrations (per-C allele beta 0.076 SD; SE: 0.013; p = 1.5 × 10-9) but not with confounders. In Mendelian randomization analyses, apoA-I was not related to risk of CAD (OR 1.13; 95%CI: 0.98,1.30 per 1-SD higher apoA-I), which was different from the observational association. Similar findings were observed using an independent ABCA1 variant in sensitivity analysis. Genetic evidence fails to support a cardioprotective role for apoA-I. This is in line with the cumulative evidence showing that HDL-related phenotypes are unlikely to have a protective role in CAD.


Changes in neurophysiologic markers of visual processing following beneficial anti-VEGF treatment in macular degeneration.
Antivascular endothelial growth factor (VEGF) agents have been shown to improve visual acuity and prevent vision loss in exudative age-related macular degeneration. As the vision improves relatively quickly in response to intravitreal injections, we wanted to know whether this improvement is reflected in electrophysiological markers of visual cortical processing. Our interventional case series included six elderly patients who underwent injection treatment to the affected eye. Their visual acuity, tomographic images of retinal thickness, and visual evoked potentials (VEP) were assessed before treatment and six weeks after the last injection. All patients showed improved visual acuity and reduced retinal fluid after the treatment. All but one patient showed increased VEP P100 component amplitudes and/or shortened latencies in the treated eye. These VEP changes were consistent with improved vision while the untreated eyes showed no changes. Our results indicate that antivascular endothelial growth factor injections improved visual function of the treated eyes both in the level of the retina and in the level of visual cortical processing.


Illness perceptions and their association with 2 year functional status and change in patients with hand osteoarthritis.
To investigate the association between illness perceptions and disability both cross-sectionally and over 2 years in patients with hand OA. Illness perceptions and self-reported disability were assessed at baseline and after 2 years in 384 patients with primary hand OA (mean age 61 years, 84% women, n = 312 with follow-up) with the Illness Perception Questionnaire - Revised (IPQ-R), Functional Index for Hand OA, Australian/Canadian Hand OA Index and HAQ. Risk ratios for high disability (highest quartile) at both time points were estimated for tertiles of IPQ-R dimensions, using Poisson regression. The mean IPQ dimension change difference between patients with and without disability progression (change Functional Index for Hand OA ⩾ 1, Australian/Canadian Hand OA Index > 1.4, HAQ > 0.22) was estimated with linear regression. Analyses were adjusted for age, Doyle index and baseline score. At baseline, stronger negative illness perceptions were associated with high disability. Baseline illness perceptions were also associated with high disability after 2 years, although adjustment made apparent that these associations were confounded by baseline disability status. Most illness perceptions changed over 2 years; understanding increased, OA was regarded as more chronic and fewer emotions and consequences and less personal and treatment control were experienced. The 2 year change in disability was different between patients with and without progression for the illness perceptions of more perceived consequences, symptoms, treatment control and emotions. Illness perceptions seemed to be implicated in disability and its progression. Our results suggest that interventions could focus on improving baseline disability, potentially using illness perceptions to accomplish this goal.


Association of hepatitis C virus and hepatocellular carcinoma in the United States.
Hepatocellular carcinoma (HCC) is a common cause of death from cancer on a worldwide basis. Chronic hepatitis B virus (HBV) infection is a well-documented pathogenic factor in the etiology of HCC, accounting for the majority of cases in certain geographic areas where HBV chronic-infection rates are high. Recent studies have implicated hepatitis C virus (HCV) infection as another major etiologic agent of hepatitis B surface antigen (HBsAg)-negative HCC. Studies in Europe have demonstrated that HCV infection is commonly associated with HCC; the prevalence rates of HCV ranged from 39% to 77% of HCC patients in these studies. In areas where HBV infection is endemic, such as Africa and Japan, the prevalence rates of anti-HCV in individuals with HCC have been reported at 29% and 68%, respectively. The significance of HCV and HBV infections in contributing to the occurrence of HBsAg-negative HCC has not been well established in the United States. The clinical consequences of infection with different genotypes of HCV (defined by nucleotide sequence divergence) are largely unknown: several studies have suggested differences among genotypes in terms of clinical expression, disease progression, and response to treatment with interferon. Our recent studies demonstrated that HCV and occult HBV infections account for the majority of cryptogenic HCC cases in the United States. Preliminary studies on the prevalence of HCV genotypes in patients with or without HCC but otherwise matched for hepatic histology did not reveal any association of specific genotype(s) with development of HCC.


Pitfalls in the statistical examination and interpretation of the correspondence between physician and patient satisfaction ratings and their relevance for shared decision making research.
The correspondence of satisfaction ratings between physicians and patients can be assessed on different dimensions. One may examine whether they differ between the two groups or focus on measures of association or agreement. The aim of our study was to evaluate methodological difficulties in calculating the correspondence between patient and physician satisfaction ratings and to show the relevance for shared decision making research. We utilised a structured tool for cardiovascular prevention (arriba™) in a pragmatic cluster-randomised controlled trial. Correspondence between patient and physician satisfaction ratings after individual primary care consultations was assessed using the Patient Participation Scale (PPS). We used the Wilcoxon signed-rank test, the marginal homogeneity test, Kendall's tau-b, weighted kappa, percentage of agreement, and the Bland-Altman method to measure differences, associations, and agreement between physicians and patients. Statistical measures signal large differences between patient and physician satisfaction ratings with more favourable ratings provided by patients and a low correspondence regardless of group allocation. Closer examination of the raw data revealed a high ceiling effect of satisfaction ratings and only slight disagreement regarding the distributions of differences between physicians' and patients' ratings. Traditional statistical measures of association and agreement are not able to capture a clinically relevant appreciation of the physician-patient relationship by both parties in skewed satisfaction ratings. Only the Bland-Altman method for assessing agreement augmented by bar charts of differences was able to indicate this. ISRCTN: ISRCT71348772.


The left-right determinant inversin has highly conserved ankyrin repeat and IQ domains and interacts with calmodulin.
All vertebrates have a left-right body axis with invariant asymmetries of the heart and the positions of the abdominal viscera. Major advances have recently been made in defining molecular components of the pathway specifying the vertebrate left-right axis, but our knowledge of the early determinants is extremely limited. In the invmouse the left-right axis is consistently reversed, unlike other vertebrate mutants where randomisation of situs is apparent. The gene disrupted in this mouse encodes a 1062-amino-acid protein, inversin. We previously reported 16 tandem ankyrin repeats, spanning amino acids 13-557, and two putative nuclear localisation sequences, but otherwise the sequence offered few clues to the possible function. In order to identify regions likely to be functionally important, we have identified and characterised orthologous sequences in several species, including chick, Xenopus and zebrafish. Sequence comparisons show strong conservation of the ankyrin repeat region and also a lysine-rich domain spanning amino acids 558-604. Further analysis identified a highly conserved IQ calmodulin-binding domain in the latter region and another such domain in an otherwise poorly conserved C-terminal region. A yeast two-hybrid screen identified calmodulin in one third of the positive clones, and we confirmed this interaction by immunoprecipitation.


Development of a 3D collagen scaffold coated with multiwalled carbon nanotubes.
Carbon nanotubes (CNTs) have attractive biochemical properties such as strong cell adhesion and protein absorption, which are very useful for a cell cultivation scaffold. In this study, we prepared a multiwalled carbon nanotube-coated collagen sponge (MWCNT-coated sponge) to improve the surface properties of the collagen sponge, and its cell culturing properties were examined. The suface of the collagen sponge was homogeneously coated with MWCNTs by dispersion. MC3T3-E1 cells were cultured on and inside the MWCNT-coated sponge. The DNA content on the MWCNT-coated sponge after 1 week of culture was significantly higher than on an uncoated collagen sponge (p < 0.05). There was no significant difference between the estimated ALP activity normalized by DNA quantity on the MWCNT-coated sponge and that on the uncoated collagen sponge which is well known as one of the best scaffolds for cell cultivation. In addition, the MWCNT-coated surface shows strong cell adhesion. Therefore, the MWCNT-coated collagen sponge is expected to be a useful 3D scaffold for cell cultivation. (c) 2009 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2009.


Total quality management in action: pharmacy system changes to decrease medication incidents and increase clinical services.
During a monitoring of risk management indicators (incident report analysis), a trend of increasing medication incidents was noted from 1987 to 1990. The medication ordering process was tracked using a flow chart, and, using an interdisciplinary team, the transcription system was targeted for corrective action. A "roving pharmacist" system was devised, with a resultant 63% decrease in the reported medication incident rate from 1990 to 1993.


Sequence relationships between plasmids carrying genes for lactose utilization.
Southern hybridization experiments carried out between pSC101::Tn951 DNA and lambda dlac DNA allowed the location and orientation of the lac operon within the transposon to be deduced. The same method was used to detect Tn951 on Lac plasmids from 11 independent isolates from three continents. None of these plasmids was found to carry an entire Tn951 sequence but they all contained lac genes homologous to the lac genes of Tn951. The lac operon of one of these plasmids was bordered by a sequence homologous to that found at the left-hand side of Tn951. It is concluded that the lac determinants of the Lac plasmids analysed and of Tn951 have evolved from a common ancestor but that the distribution of these determinants cannot be attributed to a spread of the transposon Tn951.


Phonetic inventories, 15-24 months: a longitudinal study.
Longitudinal samples of meaningful speech of 34 normally developing children were analyzed to determine the range and types of consonantal phones produced at 15, 18, 21, and 24 months. Separate inventories for word-initial and word-final consonants were constructed for each child at each age level. Group analyses showed that early inventories in initial position were composed primarily of voiced anterior stops, nasals, and glides; by 24 months, voiceless stops, velars, and a few fricatives were also included. In final position, inventories consisted primarily of voiceless stops and alveolar consonants. There was a strong tendency for the voiced stops to appear first in initial position and for [t] and [r] to appear first in word-final. Individual analyses of place and manner of articulation revealed highly similar patterns across subjects. The findings are related to other longitudinal research in early phonological development and to studies of babbling of younger subjects and correct productions of older subjects.


Allele and genotype frequencies of polymorphic cytochromes P4502D6, 2C19 and 2E1 in aborigines from western Australia.
The polymorphisms of the important xenobiotic metabolizing enzymes CYP2D6, CYP2C19 and CYP2E1 have been studied extensively in a large number of populations and show significant heterogeneity in the frequency of different alleles/genotypes and in the prevalence of the extensive and poor metabolizer phenotypes. Understanding of inter-ethnic differences in genotypes is important in prediction of either beneficial or adverse effects from therapeutic agents and other xenobiotics. Since no data were available for Australian Aborigines, we investigated the frequencies of alleles and genotypes for CYP2D6, CYP2C19 and CYP2E1 in a population living in the far north of Western Australia. Because of its geographical isolation, this population can serve as a model to study the impact of evolutionary forces on the distribution of different alleles for xenobiotic metabolizing enzymes. Twelve CYP2D6 alleles were analysed. The wild-type allele *1 was the most frequent (85.81%) and the non-functional alleles (*4, * 5, * 16) had an overall frequency of less than 10%. Only one subject (0.4%) was a poor metabolizer for CYP2D6 because of the genotype *5/*5. For CYP2C19, the frequencies of the *1 (wild-type) and the non-functional (*2 and *3) alleles were 50.2%, 35.5% and 14.3%, respectively. The combined CYP2C19 genotypes (*2/*2, *2/*3 or *3/*3) correspond to a predicted frequency of 25.6% for the CYP2C19 poor metabolizer phenotype. For CYP2EI, only one subject had the rare c2 allele giving an overall allele frequency of 0.2%. For CYP2D6 and CYP2C19, allele frequencies and predicted phenotypes differed significantly from those for Caucasians but were similar to those for Orientals indicating a close relationship to East Asian populations. Differences between Aborigines and Orientals in allele frequencies for CYP2D6* 10 and CYP2E1 c2 may have arisen through natural selection, or genetic drift, respectively.


[A case of resected liver metastasis from rectal cancer with bile duct stricture after radiofrequency ablation].
The patient was a 66-year-old woman who had undergone low anterior resection for rectal cancer. Although she received adjuvant chemotherapy after the surgery, a 1.5 cm hepatic metastasis was observed in segments 4/8. Radiofrequency ablation ( RFA) was performed for the metastatic tumor. Approximately 1 month after RFA, a local recurrence was identified together with intrahepatic bile duct dilation in the right lobe of the liver. Extended right hepatic lobectomy and left intrahepatic cholangiojejunostomy were performed to remove the tumor. Histopathological examination of the resected specimen showed a liver metastasis from the rectal cancer with stricture of the bile duct although the bile duct was not involved in the tumor. This suggested that the bile duct stricture was caused by RFA and not by the recurrent tumor. Currently, the patient is alive with no recurrence 14 months after the surgery. This case suggests the importance of salvage surgery for recurrent tumors after RFA therapy, especially in the case of RFA-related complications in conjunction with the tumor.


Assessment of the Photosynthetic Apparatus Functions by Chlorophyll Fluorescence and P700 Absorbance in C3 and C4 Plants under Physiological Conditions and under Salt Stress.
Functions of the photosynthetic apparatus of C3 (Pisum sativum L.) and C4 (Zea mays L.) plants under physiological conditions and after treatment with different NaCl concentrations (0-200 mM) were investigated using chlorophyll a fluorescence (pulse-amplitude-modulated (PAM) and JIP test) and P700 photooxidation measurement. Data revealed lower density of the photosynthetic structures (RC/CSo), larger relative size of the plastoquinone (PQ) pool (N) and higher electron transport capacity and photosynthetic rate (parameter RFd) in C4 than in C3 plants. Furthermore, the differences were observed between the two studied species in the parameters characterizing the possibility of reduction in the photosystem (PSI) end acceptors (REo/RC, REo/CSo and δRo). Data revealed that NaCl treatment caused a decrease in the density of the photosynthetic structures and relative size of the PQ pool as well as decrease in the electron transport to the PSI end electron acceptors and the probability of their reduction as well as an increase in the thermal dissipation. The effects were stronger in pea than in maize. The enhanced energy losses after high salt treatment in maize were mainly from the increase in the regulated energy losses (ΦNPQ), while in pea from the increase in non-regulated energy losses (ΦNO). The reduction in the electron transport from QA to the PSI end electron acceptors influenced PSI activity. Analysis of the P700 photooxidation and its decay kinetics revealed an influence of two PSI populations in pea after treatment with 150 mM and 200 mM NaCl, while in maize the negligible changes were registered only at 200 mM NaCl. The experimental results clearly show less salt tolerance of pea than maize.


Parental self-efficacy and online support among parents of children diagnosed with type 1 diabetes mellitus.
Families faced with the challenges of caring for a child newly diagnosed with type 1 diabetes mellitus are often overwhelmed by the regimented demands of diabetes care management. Social support for families has been recognized as an important component to adaptation and has been deemed necessary for helping families develop healthy coping strategies. The purpose of this evidence-based practice (EBP) project was to develop a Web-based platform to enhance social support and increase self-efficacy of parents with a child with type 1 diabetes. The following clinical question guided the project: "Among parents of children diagnosed with type 1 diabetes in a small Midwestern city, what are parents' self-reported self-efficacy scores related to diabetic care management pre- and post-implementation of a Web-based social support platform?" A one-group pretest/post-test descriptive design was used, with parental self-efficacy measured pre- and post-intervention (Web-based platform access) using the Diabetes Empowerment Scale (DES) and Self-Efficacy for Diabetes Scale (SED), and parent satisfaction with the online support platform assessed through parental comments. Project outcomes corroborated the use of online social support as evidenced by improvement in parental self-efficacy scores in both the DES and SED survey measurements. A statistically significant increase (p < 0.05) was found between the pre- and post-implementation scores of the SED survey. Secondary data supported the positive relationship of social support and self-efficacy in raising a child with type 1 diabetes. This online social support platform was found to be an easily adaptable, cost-effective, and innovative means of networking and information sharing among families facing similar challenges in raising a child with type 1 diabetes.


Molecular simulation of thin liquid films: Thermal fluctuations and instability.
The instability of a thin liquid film on a solid surface is studied both by molecular dynamics simulations (MD) and a stochastic thin-film equation (STF), which models thermal fluctuations with white noise. A linear stability analysis of the STF allows us to derive a power spectrum for the surface fluctuations, which is quantitatively validated against the spectrum observed in MD. Thermal fluctuations are shown to be critical to the dynamics of nanoscale films. Compared to the classical instability mechanism, which is driven by disjoining pressure, fluctuations (a) can massively amplify the instability, (b) cause the fluctuation wavelength that is dominant to evolve in time (a single fastest-growing mode does not exist), and (c) decrease the critical wavelength so that classically stable films can be ruptured.


Comparison of Fasting Insulin Level, Homeostatic Model of Insulin Resistance, and Lipid Levels between Patients with Primary Hypertension and Normotensive Subjects.
Hyperinsulinemia and insulin resistance occurs in obese patients with primary hypertension independent of diabetes and obesity. This study was aimed at assessing serum fasting insulin levels, the homeostatic model assessment for insulin resistance (HOMA-IR), and serum lipid levels in non-obese patients with primary hypertension when compared to normotensive subjects. This observational study comprised 100 patients over 18 years of age, divided into two groups. The hypertensive group comprised non-obese patients with primary hypertension (n=50); the normotensive group comprised normotensive age- and sex-matched individuals (n=50). Patients with diabetes, impaired fasting glucose, obesity, and other causative factors of insulin resistance were excluded from the study. Serum fasting insulin levels and fasting lipid profiles were measured, and insulin resistance was calculated using HOMA-IR. These data were compared between the two groups. Pearson's correlation coefficient was used to assess the extent of a linear relationship between HOMA-IR and to evaluate the association between HOMA-IR and systolic and diastolic blood pressures. Mean serum fasting insulin levels (mIU/L), mean HOMA-IR values, and fasting triglyceride levels (mg/dL) were significantly higher in the hypertensive versus normotensive patients (10.32 versus 6.46, P<0.001; 1.35 versus 0.84, P<0.001; 113.70 versus 97.04, P=0.005, respectively). The HOMA-IR levels were associated with systolic blood pressure (r value 0.764, P=0.0005). We observed significantly higher fasting insulin levels, serum triglyceride levels, and HOMA-IR reflecting hyperinsulinemia and possibly an insulin-resistant state among primary hypertension patients with no other causally linked factors for insulin resistance. We observed a significant correlation between systolic blood pressure and HOMA-IR.


Towards a dynamical network view of brain ischemia and reperfusion. Part I: background and preliminaries.
The general failure of neuroprotectants in clinical trials of ischemic stroke points to the possibility of a fundamental blind spot in the current conception of ischemic brain injury, the "ischemic cascade". This is the first in a series of four papers whose purpose is to work towards a revision of the concept of brain ischemia by applying network concepts to develop a bistable model of brain ischemia. This first paper sets the stage for developing the bistable model of brain ischemia. Necessary background in network theory is introduced using examples from developmental biology which, perhaps surprisingly, can be adapted to brain ischemia with only minor modification. Then, to move towards a network model, we extract two core generalizations about brain ischemia from the mass of empirical data. First, we conclude that all changes induced in the brain by ischemia can be classified as either damage mechanisms that contribute to cell death, or stress responses that contribute to cell survival. Second, we move towards formalizing the idea of the "amount of ischemia", I, as a continuous, nonnegative, monotonically increasing quantity. These two generalizations are necessary precursors to reformulating brain ischemia as a bistable network.


Improving the collection efficiency of the liquid impinger for ultrafine particles and viral aerosols by applying granular bed filtration.
Liquid impingers are utilized to collect bioaerosols for many advantages, such as avoiding dehydration of biological agents. However, many previous studies have reported that the liquid impingers are surprisingly inefficient for the collection of ultrafine bioaerosols, with collection efficiencies <30%. In the present work, we have successfully improved the collection efficiency of the liquid impinger (AGI30) to as high as 99% for particles in the size range of 20-400 nm with the aid of packed glass beads. We also systematically investigated the effects of influential factors on the collection efficiency. These factors include the volume of the sampling liquid (0, 20 and 30 mL), depth (0, 7 and 10 cm) of packed glass beads and sampling flow rate (4, 6 and 8 liter per min, lpm). According to our experimental results, increasing the depth of packed glass beads and the volume of sampling liquid can enhance the collection efficiency. Also, decreasing the sampling flow rate can increase the collection efficiency and reduce the loss of sampling liquid. For the sampling of viable MS2 phages, the collection efficiency of AGI30 sampler with packed glass beads is much higher than that without packed glass beads. Conclusively, this study validates that the granular bed filtration can enhance the collection efficiency of liquid impingers for submicron and ultrafine particles and viral aerosols.


Fabrication of pH-Sensitive Tetramycin Releasing Gel and Its Antibacterial Bioactivity against Ralstonia solanacearum.
Ralstonia solanacearum (R. solanacearum)-induced bacterial wilt of the nightshade family causes a great loss in agricultural production annually. Although there has been some efficient pesticides against R. solanacearum, inaccurate pesticide releasing according to the onset time of bacterial wilt during the use of pesticides still hinders the disease management efficiency. Herein, on the basis of the soil pH change during R. solanacearum growth, and pH sensitivity of the Schiff base structure, a pH-sensitive oxidized alginate-based double-crosslinked gel was fabricated as a pesticide carrier. The gel was prepared by crosslinking oxidized sodium alginate (OSA) via adipic dihydrazide (ADH) and Ca2+. After loading tetramycin into the gel, it showed a pH-dependent pesticide releasing behavior and anti-bacterial activity against R. solanacearum. Further study also showed that the inhibition rate of the tetramycin-loaded gel was higher than that of industrial pesticide difenoconazole. This work aimed to reduce the difficulty of pesticide administration in the high incidence period of bacterial wilt and we believe it has a great application potential in nightshade production.


Endothelial and metabolic characteristics of patients with angina and angiographically normal coronary arteries: comparison with subjects with insulin resistance syndrome and normal controls.
This study was performed to characterize the endothelial and metabolic alterations of patients with angina and angiographically normal coronary arteries ("cardiac" syndrome X [CSX]) compared with subjects with insulin resistance syndrome ("metabolic" syndrome X [MSX]) and normal controls. Previous studies have found high endothelin-1 levels, impaired endothelium-dependent vasodilation and insulin resistance in patients with angina pectoris and angiographically normal coronary arteries. On the other hand, subjects with insulin resistance syndrome have shown high endothelin-1 levels. Thirty-five subjects were studied: 13 patients with angina pectoris and angiographically normal coronary arteries (CSX group); 9 subjects with insulin resistance syndrome (MSX group) and 13 normal controls. All subjects received an acute intravenous bolus of insulin (0.1 U/kg) combined with a euglycemic clamp and forearm indirect calorimetry. Endothelin-1 levels, nitrite/nitrate (NOx) levels, end products of nitric oxide metabolism, glucose infusion rates (index of insulin sensitivity) and their incremental areas (deltaAUCs [area under curves]) were measured during this period. Basal endothelin-1 levels were higher in CSX and MSX groups than in normal controls (8.19 +/- 0.46 and 6.97 +/- 0.88 vs. 3.67 +/- 0.99 pg/ml; p < 0.01), while basal NOx levels were significantly higher in MSX group than in CSX and normal controls (36.5 +/- 4.0 vs. 24.2 +/- 3.3 and 26.8 +/- 3.2 mol/liter, p < 0.05). After insulin administration, the deltaAUCs of NOx (p < 0.05) were lower in CSX group than in MSX and normal controls, and the deltaAUCs of endothelin-1 were lower in group CSX than in normal controls. Glucose infusion rate was significantly lower in CSX and MSx groups than in normal controls (p < 0.01), suggesting that in both CSX and MSX groups insulin resistance is present. A positive correlation was found between the deltaAUCs of nitric oxide and the AUCs of glucose infusion rate. Blunted nitric oxide and endothelin responsiveness to intravenously infused insulin is a typical feature of patients with angina pectoris and angiographically normal coronary arteries and may contribute to the microvascular dysfunction observed in these subjects.


Usability of a Co-designed eHealth Prototype for Caregivers: Combination Study of Three Frameworks.
Co-design (or the participation of users) has shown great potential in the eHealth domain, demonstrating positive results. Nevertheless, the co-design approach cannot guarantee the usability of the system designed, and usability assessment is a complex analysis to perform, as evaluation criteria will differ depending on the usability framework (or set of criteria) used. ISO (International Organization for Standardization) on usability (ISO 9241-210), Nielsen heuristic, and Garrett element of user experience inform different yet complementary aspects of usability. This study aims to assess the usability and user experience of a co-design prototype by combining 3 complementary frameworks. To help caregivers provide care for functionally impaired older people, an eHealth tool was co-designed with caregivers, health and social service professionals, and community workers assisting caregivers. The prototype was a website that aims to support the help-seeking process for caregivers (finding resources) and allow service providers to advertise their services (offering resources). We chose an exploratory study method to assess usability in terms of each objective. The first step was to assess users' first impressions of the website. The second was a task scenario with a think-aloud protocol. The final step was a semistructured interview. All steps were performed individually (with a moderator) in a single session. The data were analyzed using 3 frameworks. A total of 10 participants were recruited, 5 for each objective of the website. We were able to identify several usability problems, most of which were located in the information design and interface design dimensions (Garrett framework). Problems in both dimensions were mainly coded as effectiveness and efficiency (ISO framework) and error prevention and match between the systemand the real world (Nielsen heuristic). Our study provided a novel contribution about usability analysis by combining the 3 different models to classify the problems found. This combination provided a holistic understanding of the usability improvements needed. It can also be used to analyze other eHealth products. RR2-10.2196/11634.


MiR-802 Suppresses Colorectal Cancer Cell Viability, Migration and Invasion by Targeting RAN.
Colorectal cancer is one of the most malignant tumors in the world, and the incidence is increasing every year. MicroRNAs (miRNA) are small non-coding RNAs that are involved in a variety of physiological or pathological processes. Abnormal expression of microRNA-802 (miR-802) has been demonstrated in various types of cancer. However, the expression and biological role of miR-802 in human colorectal cancer remain largely unknown. Here, we used quantitative real-time PCR (qRT-PCR) to measure miR-802 expression levels in colorectal cancer tissues and cell lines. Cell Counting Kit-8 (CCK-8) was used to assess the effect of miR-802 on colorectal cancer cell viability. Migration and invasion assays were performed to determine the effect of miR-802 on metastasis of colon tumor cells by transwell analysis. Luciferase activity assays were used to confirm the target of miR-802. The results show that miR-802 is significantly downregulated in colorectal cancer tissues and cell lines. Overexpression of miR-802 profoundly inhibited viability, migration and invasion of colorectal cancer cells. In addition, we have newly discovered that the Ras-associated nucleus (RAN) is a direct target of miR-802 which could reverse the effects induced by miR-802 overexpression in colorectal cancer cells. In conclusion, our study shows that miR-802 is downregulated in colorectal cancer, and overexpression of miR-802 inhibits colorectal cancer cell viability, migration and invasion by directly targeting RAN.


BR96-DOX immunoconjugate targeting of chemotherapy in brain tumor models.
An immunoconjugate of doxorubicin (adriamycin) and a tumor-specific monoclonal antibody, BR96-DOX (now SGN-15) targets chemotherapy to cells that express the LewisY antigen. This immunoconjugate is internalized into lysosomes in antigen-expressing cells, with release of free doxorubicin after hydrolysis of the acid-labile linker. We review our studies using BR96-DOX in a human small-cell lung carcinoma intracerebral xenograft model in nude rats. We have found that the immunoconjugate is effective against intracerebral tumors when delivery is enhanced with osmotic disruption of the blood-brain barrier (BBB). Enhanced delivery of BR96-DOX with BBB opening can work together with radiotherapy to increase antitumor efficacy, which is maximally effective if immunoconjugate is administered prior to radiotherapy. In heterogeneous brain tumors, enhanced delivery of BR96-DOX significantly reduced tumor volumes, but local release of doxorubicin by targeting antigen expressing cells shows modest cytotoxicity against adjacent non-expressor cells. Although BR96-DOX is not effective against glioma cells tested, it does provide a model for drug-immunoconjugate therapy of gliomas. Our studies in a rat brain tumor model point out the importance of optimized delivery, antigenic heterogeneity, and bystander effect for brain tumor therapy. We review additional studies of drug-mAb immunoconjugates pertinent to the treatment of gliomas.


Biodistribution of Biomimetic Drug Carriers, Mononuclear Cells, and Extracellular Vesicles, in Nonhuman Primates.
Discovery of novel drug delivery systems to the brain remains a key task for successful treatment of neurodegenerative disorders. Herein, the biodistribution of immunocyte-based carriers, peripheral blood mononuclear cells (PBMCs), and monocyte-derived EVs are investigated in adult rhesus macaques using longitudinal PET/MRI imaging. 64 Cu-labeled drug carriers are introduced via different routes of administration: intraperitoneal (IP), intravenous (IV), or intrathecal (IT) injection. Whole body PET/MRI (or PET/CT) images are acquired at 1, 24, and 48 h post injection of 64 Cu-labeled drug carriers, and standardized uptake values (SUVmean and SUVmax ) in the main organs are estimated. The brain retention for both types of carriers increases based on route of administration: IP < IV < IT. Importantly, a single IT injection of PBMCs produces higher brain retention compared to IT injection of EVs. In contrast, EVs show superior brain accumulation compared to the cells when administered via IP and IV routes, respectively. Finally, a comprehensive chemistry panel of blood samples demonstrates no cytotoxic effects of either carrier. Overall, living cells and EVs have a great potential to be used for drug delivery to the brain. When identifying the ideal drug carrier, the route of administration could make big differences in CNS drug delivery.


N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline, amiloride analogues, and renal Na+/H+ antiporter.
N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ) is a carboxyl-activating agent and has been shown to inhibit the renal Na+/H+ antiporter. The purposes of the present studies were to characterize the kinetics of inhibition of the Na+/H+ antiporter by EEDQ and to determine whether amiloride analogues affect the ability of EEDQ to inhibit the rate of Na+/H+ exchange. Brush-border membrane vesicles (BBMV) were prepared from rabbit kidneys; Na+/H+ exchange rate was assessed by the fluorescence quenching of acridine orange. EEDQ produced a concentration-dependent inhibition of Na+/H+ exchange; the effect was to decrease the maximum activity (Vmax) from 5.51 to 2.03 fluorescence units X mg protein-1 X S-1) and Km (from 14.1 to 8.7 mM) compared with control BBMV. Pretreatment of BBMV with amiloride before the addition of EEDQ maintained both Vmax and Km at values that were not significantly different from those for control BBMV. Compared with a series of analogues, amiloride was only the third most potent inhibitor of the rabbit renal Na+/H+ antiporter; amiloride, however, provided the greatest protection against inhibition of the antiporter by the subsequent addition of EEDQ. These findings suggest that the 2-carbonylguanidininum moiety and 6-chloro atom are important for binding of amiloride to sites at or near the antiporter; the group at position 5 is important in determining the ability of amiloride to protect against inhibition of the Na+/H+ antiporter by EEDQ. Finally, the ability of amiloride to protect against inactivation of the renal Na+/H+ antiporter by EEDQ is reversible.


Shedding of OXA-181 carbapenemase-producing Escherichia coli from companion animals after hospitalisation in Switzerland: an outbreak in 2018.
BackgroundCarbapenem-resistant Enterobacteriaceae pose a serious threat to public health worldwide, and the role of companion animals as a reservoir is still unclear.AimsThis 4-month prospective observational study evaluated carriage of carbapenem-resistant Enterobacteriaceae at admission and after hospitalisation in a large referral hospital for companion animals in Switzerland.MethodsRectal swabs of dogs and cats expected to be hospitalised for at least 48 h were taken from May to August 2018 and analysed for the presence of carbapenem-resistant Enterobacteriaceae using selective agar plates. Resistant isolates were further characterised analysing whole genome sequences for resistance gene and plasmid identification, and ad hoc core genome multilocus sequence typing.ResultsThis study revealed nosocomial acquisition of Escherichia coli harbouring the carbapenemase gene bla OXA-181, the pAmpC cephalosporinase gene bla CMY-42 as well as quinolone resistance associated with qnrS1 and mutations in the topoisomerases II (GyrA) and IV (ParC). The bla OXA-181 and qnrS1 genes were identified on a 51 kb IncX3 plasmid and bla CMY-42 on a 47 kb IncI1 plasmid. All isolates belonged to sequence type ST410 and were genetically highly related. This E. coli clone was detected in 17 of 100 dogs and four of 34 cats after hospitalisation (21.6%), only one of the tested animals having tested positive at admission (0.75%). Two positive animals were still carriers 4 months after hospital discharge, but were negative after 6 months.ConclusionsCompanion animals may acquire carbapenemase-producing E. coli during hospitalisation, posing the risk of further dissemination to the animal and human population and to the environment.


[The ultrastructure of the white rat olfactory epithelium].
As a result of electron microscope observation of olfactory mucosa in the white rat, variability of structure of the supporting cells was established. Among these cells secretory ones and those rich in endoplasmic reticulum were discovered. The differences revealed in dendrites and rods of olfactory cells seem to be associated with functional condition of these cells.


Comparative study of the dental substrate used in shear bond strength tests.
The purpose of this study was to compare shear bond strength values obtained in human enamel and dentin with the values obtained in bovine teeth using two adhesive systems with different actions. Forty human tooth half-crowns and forty bovine tooth crowns were flattened to a minimum plain area of 5 mm in diameter. The samples were divided in four groups of 20 specimens each: 1) human enamel; 2) bovine enamel; 3) human dentin; 4) bovine dentin. The samples of each group were divided in 2 subgroups of 10 samples each, according to the adhesive system used: 1) Scotchbond Multi-Purpose (SBMP); and 2) Clearfil Liner Bond 2V (CLB2V) applied according to the manufacturer's recommendations. Afterwards, restorations of Z100 composite with cylindrical shape (4 mm diameter x 5 mm height) were made using a metallic mold to submit the samples to shear bond testing on an Instron universal testing machine, at a crosshead speed of 0.5 mm/min. The data were submitted to ANOVA and Tukey's test (5%). In enamel, there was no statistical difference between bovine and human teeth for SBMP (7.36 MPa, human; 8.24 MPa, bovine), nor for CLB2V (10.01 MPa, human; 7.95, bovine). In dentin, SBMP showed a statistically lower mean on human dentin (7.01 MPa) than on bovine dentin (11.74 MPa). For CLB2V, there was no statistical difference between human (7.43 MPa) and bovine (9.27 MPa) substrates.


Quantitative assessment of lung stiffness in patients with interstitial lung disease using MR elastography.
To investigate the use of magnetic resonance elastography (MRE) in the quantitative assessment of pulmonary fibrosis by comparing quantitative shear stiffness measurements of lung parenchyma in patients diagnosed with fibrotic interstitial lung disease (ILD) and healthy controls. A 1.5T spin-echo, echo planar imaging MRE (SE-EPI MRE) pulse sequence was utilized to assess absolute lung shear stiffness in 15 patients with diagnosed ILD and in 11 healthy controls. Data were collected at residual volume (RV) and total lung capacity (TLC). Spirometry data were obtained immediately prior to scanning. To test for statistical significance between RV and TLC shear stiffness estimates a two-sample t-test was performed. To assess variability within individual subject shear stiffness estimates, the intraclass correlation coefficient (ICC) and Krippendorff's alpha were calculated. Patients with ILD exhibited an average (±1 standard deviation) shear stiffness of 2.74 (±0.896) kPa at TLC and 1.32 (±0.300) kPa at RV. The corresponding values for healthy individuals were 1.33 (±0.195) kPa and 0.849 (±0.250) kPa, respectively. The difference in shear stiffness between RV and TLC was statistically significant (P < 0.001). At TLC, the ICC and alpha values were 0.909 and 0.887, respectively. At RV, the ICC and alpha values were 0.852 and 0.862, respectively. In subjects with known fibrotic interstitial lung disease, parenchymal shear stiffness is increased when compared to normal controls at both RV and TLC, with TLC demonstrating the most significant difference. MRE-derived parenchymal shear stiffness is a promising new noninvasive imaging-based biomarker of interstitial lung disease. 1 Technical Efficacy: Stage 2 J. MAGN. RESON. IMAGING 2017;46:365-374.


Interaction between the basolateral K+ and apical Na+ conductances in Necturus urinary bladder.
Experimental modulation of the apical membrane Na+ conductance or basolateral membrane Na+-K+ pump activity has been shown to result in parallel changes in the basolateral K+ conductance in a number of epithelia. To determine whether modulation of the basolateral K+ conductance would result in parallel changes in apical Na+ conductance and basolateral pump activity, Necturus urinary bladders stripped of serosal muscle and connective tissue were impaled through their basolateral membranes with microelectrodes in experiments that allowed rapid serosal solution changes. Exposure of the basolateral membrane to the K+ channel blockers Ba2+ (0.5 mM/liter), Cs+ (10 mM/liter), or Rb+ (10 mM/liter) increased the basolateral resistance (Rb) by greater than 75% in each case. The increases in Rb were accompanied simultaneously by significant increases in apical resistance (Ra) of greater than 20% and decreases in transepithelial Na+ transport. The increases in Ra, measured as slope resistances, cannot be attributed to nonlinearity of the I-V relationship of the apical membrane, since the measured cell membrane potentials with the K+ channel blockers present were not significantly different from those resulting from increasing serosal K+, a maneuver that did not affect Ra. Thus, blocking the K+ conductance causes a reduction in net Na+ transport by reducing K+ exit from the cell and simultaneously reducing Na+ entry into the cell. Close correlations between the calculated short-circuit current and the apical and basolateral conductances were preserved after the basolateral K+ conductance pathways had been blocked. Thus, the interaction between the basolateral and apical conductances revealed by blocking the basolateral K+ channels is part of a network of feedback relationships that normally serves to maintain cellular homeostasis during changes in the rate of transepithelial Na+ transport.


Identification of active sites in CO oxidation over a Pd/Al2O3 catalyst.
The active sites of Pd/Al2O3 catalysts for CO oxidations were identified by investigating the dependence of CO oxidation activities on the surface structure and morphology of Pd nanoparticles. The maximum catalytic activity was obtained for Pd particles approximately 2 nm in particle size. We performed structural analyses on the Pd surface through infrared (IR) spectroscopy of the adsorbed CO molecules. A positive correlation was obtained between catalytic activity and the fraction of linear CO adsorbed on Pd corner sites and Pd(111) facets, indicating that these sites are highly active for CO oxidation. X-ray absorption fine structure (XAFS) and spherical aberration-corrected scanning transmission electron microscopy (Cs-STEM) measurements demonstrated that Pd nanoparticles less than 2 nm in particle size with amorphous-like structures and Pd particles with large, well-ordered structures favor the formation of a high fraction of corner sites and Pd(111) facets, respectively.


[Histogenesis and structural organization of the walls of rat venae cavae and pulmonary veins].
Using light and electron microscopic methods, the histogenesis and structural organization of the walls of rat venae cavae and pulmonary veins were studied in prenatal and postnatal periods of development. The special attention was paid to the appearance of the striated myocytes in the walls of these vessels during the process of ontogenesis. The time of initial divergent development of myoblastic differon was established, the stages of differentiation of striated myoblasts and the peculiarities of intercellular junctions were characterized, as well as the innervation and vascularization of the walls of venae cavae and pulmonary veins.


Relationship between cell surface composition of Candida albicans and adherence to acrylic after growth on different carbon sources.
The adherence of Candida albicans to acrylic was measured in vitro after growth of the yeast to stationary phase in defined medium containing glucose, sucrose, galactose, fructose, or maltose as the carbon source. In each case, yeast adherence was proportional to the concentration of sugar in the growth medium, but equimolar concentrations of different sugars promoted adherence to different extents. In vitro adherence was further increased by the addition of divalent cations to assay mixtures but was inhibited when saliva-treated acrylic strips were used or when yeasts were suspended in mixed saliva during the assay. The rate of spheroplast formation of yeasts grown in media containing a 500 mM concentration of the different sugars correlated well with the relative adherence of the cells to acrylic. Galactose-grown yeasts were most resistant to spheroplast formation with Zymolyase-5000 and most adherent to acrylic, whereas fructose-grown organisms were least resistant to spheroplast formation and least adherent to acrylic. These results indicate that when grown to stationary phase in media containing high concentrations of certain sugars, C. albicans undergoes a change in cell surface composition which facilitates its adherence to acrylic surfaces. Electron microscopy of yeasts harvested from such media revealed the presence of an additional surface layer which may be responsible for this enhanced adherence.


[Phase action of ouabain on the electrical and contractile activity of the frog myocardium].
Ouabain effects (5 X 10(-8)-5 X 10(-5) M) on the isometric contraction and intracellular action potentials were studied on frog atrial and ventricular muscles. It was found that ouabain produced a three-phase action on the electrical and contractile activity of the myocardium under different patterns of stimulation both in normal Ringer and D-600-containing solutions. The data suggest that the three-phase ouabain action is a consequence of its influence on at least two component processes of the excitation-contraction coupling in the frog heart muscle: the slow Na-Ca channels of the sarcolemma and Na+-K+ pump. The contribution of each mechanism depends on the glycoside concentration and the duration of exposure to ouabain.


Clinical application and reliability of a post abdominal surgery pain assessment scale (PASPAS) in horses.
The aim of this study was to refine a multi-dimensional scale based on physiological and behavioural parameters, known as the post abdominal surgery pain assessment scale (PASPAS), to quantify pain after laparotomy in horses. After a short introduction, eight observers used the scale to assess eight horses at multiple time points after laparotomy. In addition, a single observer was used to test the correlation of each parameter with the total pain index in 34 patients, and the effect of general anaesthesia on PASPAS was investigated in a control group of eight horses. Inter-observer variability was low (coefficient of variation 0.3), which indicated good reliability of PASPAS. The correlation of individual parameters with the total pain index differed between parameters. PASPAS, which was not influenced by general anaesthesia, was a useful tool to evaluate pain in horses after abdominal surgery and may also be useful to investigate analgesic protocols or for teaching purposes.


Methicillin-resistant Staphylococcus aureus in Zimbabwe.
The prevalence of Methicillin Resistant Staphylococcus aureus (MRSA) in Africa is sparsely documented. In Zimbabwe there is no routine patient or specimen screening for MRSA. The aim of this study was to document the presence and epidemiology of MRSA in Zimbabwe. The study was done in one private sector laboratory with a national network that serves both public and private hospitals. The sample population included in-patients and outpatients, all ages, both genders, all races and only one positive specimen per patient was counted. Specimens testing positive for Staphylococcus aureus in this laboratory were further tested for MRSA using cefoxitin, by standard laboratory procedures. Data was collected from 1(st) June 2013 to 31(st) May 2014. MRSA was positive in 30 of 407 [7.0%] cases of Stapylococcus aureus reported from the laboratory. All age groups were affected from neonates to geriatrics. All specimens had similar antibiotic susceptibility pattern. Resistance was high for most widely used drugs in Zimbabwe with high sensitivity to vancomycin, linezolid and teicoplanin. Although there are no recent reports in the literature of the presence of MRSA in Zimbabwe, this study documented a 7.0% prevalence. Resistance to common antibiotics is high and antibiotic oversight is required to control the emergence of resistance to these few expensive drugs. Study was supported by Department of Anaesthesia and Critical Care funds.


Virtual Reality Meditation Among Youth Experiencing Homelessness: Pilot Randomized Controlled Trial of Feasibility.
Homelessness among youth is devastating, with high rates of substance use disorders and mental health comorbidity. Mindfulness-based interventions that include meditation and mindfulness skills training reduce stress and symptoms of anxiety or depression. However, engaging high-risk youth in interventions is challenging. Virtual reality is a more flexible platform for delivering meditation and may be appealing to youth. The study objectives were to evaluate the feasibility of delivering virtual reality meditation and of collecting outcome measures, including anxiety and physiologic stress. A sample of 30 youth experiencing homelessness was enrolled in the study. Youth were randomized to receive 10 minutes of one of three interventions: (1) virtual reality meditation, (2) audio meditation (through a web-based platform), or (3) virtual reality imagery of historical pictures and text. Subjects who consented to the study attended two research visits. The first visit collected survey measures of demographics, mental health, and substance use, and oriented subjects to the intervention platforms. The second visit (1-3 days later) delivered the intervention and collected pre and post outcome measures of anxiety and physiologic stress (salivary cortisol). Changes in anxiety and cortisol at the second visit were compared across groups using a linear regression model in which the primary analysis compared virtual reality meditation to audio meditation and secondary analyses compared virtual reality meditation to virtual reality imagery. Anxiety scores decreased in all groups, with a larger reduction among the virtual reality meditation group (difference=10.8) compared to the web-based meditation or virtual reality images groups (difference=5.8 and 5.0, respectively). After controlling for baseline values, there were no significant group differences in changes in anxiety scores or cortisol levels. In comparing virtual reality meditation and audio meditation, the effect size for anxiety was moderate (Cohen d=0.58) while the effect size for cortisol was small (Cohen d=0.08). Preliminary results suggest that virtual reality meditation has a moderate benefit for anxiety but not physiologic stress. Future research is needed to confirm these results in a larger sample and to investigate whether the effects are sustained or increase with repeated use of virtual reality mediation. Virtual reality meditation appears feasible to deliver among homeless youth and merits further study. ClinicalTrials.gov NCT04004520; https://clinicaltrials.gov/ct2/show/NCT04004520.


Nephropathy associated with Wilms' tumor. A case of a 13-year-old girl.
The association of nephropathy with Wilms' tumor and male pseudohermaphroditism is known as Drash's syndrome. The absence of either pseudohermaphroditism or Wilms' tumor has also been reported. The nephropathy, characterized by proteinuria or nephrotic syndrome, leads rapidly and inevitably to renal failure. The renal findings are those of a chronic glomerulopathy with mesangial hypercellularity, glomerular sclerosis, interstitial infiltration and marked tubular dilation. This infrequent syndrome usually appears in early life, between 1 and 3 years of age. The case of nephropathy associated with Wilms' tumor we present here is characterized by age and onset: an abrupt onset of renal insufficiency without previous signs of nephropathy in a 13-year-old girl.


Increased expression of p16 in both oral and genital lichen planus.
Lichen Planus, LP, is an inflammatory disease of possible autoimmune origin affecting mainly oral and genital mucosa and skin. According to the WHO oral LP is considered a potentially malignant disorders. The p16 tumour suppressor protein can act as an inhibitor of cyclin dependent kinases 4 and 6 and thus down regulate cell cycle progression. Since the discovery of p16 several studies have evaluated its expression in various forms of human cancers. The aim of this study was to evaluate and compare the expression of p16 in oral and genital LP and corresponding healthy mucosa. A total of 76 cases of oral LP (OLP), 34 cases of genital LP (GLP), 12 cases of healthy oral and 9 cases of healthy genital mucosa were analysed by the use of immunohistochemistry. Data showed p16 to be highly expressed in both oral and genital LP, higher than in oral (p=0.000), and genital controls (p=0.002). Results suggest that the over-expression of p16 seen in LP play a part in the histopathology of the disease.


Is There a Link Between Carotid Atherosclerosis and Idiopathic Overactive Bladder Among Women with Metabolic Syndrome?
To evaluate the association between increased intima-media thickness (IMT) and atherosclerotic carotid plaque (CP) with idiopathic overactive bladder syndrome (OAB) in women with metabolic syndrome (MetS). This is a cross-sectional study, which included consecutive women aged 40-75 years with MetS, seeking medical assistance at a reference center between April and December 2016. OAB-V8 questionnaire was used to estimate the prevalence of OAB symptoms, which were defined by a score ≥8 points. All patients underwent bilateral carotid artery ultrasound to assess IMT and CP. Atherosclerosis was defined by the identification of CP during ultrasound, which was diagnosed according to the Mannheim carotid intima-media thickness and plaque consensus. Forty-five women were prospectively included. Mean age was 60±9.3 years (range 40-75 ys). Eighteen (40%) patients were diagnosed with OAB. IMT in the general population was 0.72 mm (SD = 0.20). Overall prevalence of atherosclerosis, defined by the presence of the carotid artery plaque, was 51%. OAB prevalence among women with atherosclerosis was higher than in those without atherosclerosis (56.52% versus 22.73%), with a prevalence ratio of 2.49 (p=0.04). Additionally, OAB was associated with degree of carotid stenosis (p = 0.029). In this cohort of female patients with MetS, there was an association between carotid atherosclerosis and OAB. Identification of carotid ultrasound abnormalities may lead to refined treatment decision-making among OAB patients.


[Clinicopathological characteristics, MSI and K-ras gene mutations of double primary malignancies associated with colorectal cancer].
Objective: To investigate the clinicopathological characteristics, MSI and K-ras mutation of double primary malignancies (DPM) associated with colorectal cancer (CRC). Methods: From January 2015 to December 2016, the clinicopathological data of CRC patients treated by surgery in the Affiliated Drum Tower Hospital of Nanjing University Medical School were collected, and the clinical data was analyzed. Multiplex real-time fluorescence quantitative PCR and amplification refractory mutation was performed to identify MSI and K-ras gene mutations. Results: Of all patients with CRC, 5.2% (55/1 066) were DPM. There was no significant difference in the male and female ratio, age, colorectal cancer site, T stage, N stage composition ratio between DPM patients with CRC and patients with single CRC (P>0.05). There were significant difference of TNM stage between the two group (P<0.05). The most frequent location of CRC was the colon in both DPM patients with CRC and patients with single CRC[35.5% (359/1 011) and 41.8% (23/55), respectively]. Of 55 DPM patients with CRC, 48 were metachronous DPM patients, 7 were synchronous DPM patients and 41 were colorectal cancer first. In extracolonic organ, digestive system (23/55) was the most commonly occurring system and stomach (11/55) was the most common lesion. DPM patients with CRC had higher incidence of MSI-H than patients with single CRC (P<0.05). There was no significant difference of K-ras gene mutation between DPM patients with CRC and patients with single CRC (P>0.05). MSI-H and K-ras mutation were present in only 2 patients of DPM patients with CRC. Conclusions: The rectum is the most common lesion site in CRC patients. The stomach is the most common extracolonic organ of DPM patients with CRC. DPM patients with CRC has high risk of MSI-H, but no significant difference in the incidence of K-ras mutation.


Phospholipid synthesis in HeLa cells exposed to immunoglobulin G and complement.
1. HeLa cells were cultured in the presence of heterologous immunoglobulin G and guinea-pig serum together with [(32)P]phosphate. 2. Incorporation of [(32)P]phosphate was significantly stimulated by anti-HeLa immunoglobulin G and complement-sufficient serum compared with immunoglobulin G from unimmunized rabbits and complement. Within 2.5h heat-inactivated guinea-pig serum and anti-HeLa immunoglobulin G stimulated [(32)P]phosphate incorporation to the same extent as heat-inactivated complement and immunoglobulin G from unimmunized rabbits. 3. Compared with cells exposed to immunoglobulin G from unimmunized rabbits together with complement, anti-HeLa immunoglobulin G with complement increased the phospholipid content of HeLa cells twofold within 5h of incubation. 4. Exposure of HeLa cells to anti-HeLa immunoglobulin G and complement for 5-22h resulted in a twofold increase in the net accumulation of [(32)P]phosphate in sphingomyelin and phosphatidylcholine and a 50% increase in the net accumulation of [(32)P]phosphate in phosphatidylethanolamine, compared with cultures exposed to immunoglobulin G from unimmunized rabbits and complement. 5. A transient accumulation of (32)P-labelled lysophosphoglycerides in HeLa cells exposed to antibody and complement was detected, confirming previous findings (Güttler & Clausen, 1969b). 6. The stimulation of [(32)P]phosphate turnover occurred in cells filling up their cytoplasma with vacuoles. This supports the suggestion that the accumulation of phospholipid in these cells may be concerned with the synthesis and function of cytomembranes.


Connexin40 messenger ribonucleic acid is positively regulated by thyroid hormone (TH) acting in cardiac atria via the TH receptor.
Thyroid hormone (TH) regulates many cardiac genes via nuclear thyroid receptors, and hyperthyroidism is frequently associated with atrial fibrillation. Electrical activity propagation in myocardium depends on the transfer of current at gap junctions, and connexins (Cxs) 40 and 43 are the predominant junction proteins. In mice, Cx40, the main Cx involved in atrial conduction, is restricted to the atria and fibers of the conduction system, which also express Cx43. We studied cardiac expression of Cx40 and Cx43 in conjunction with electrocardiogram studies in mice overexpressing the dominant negative mutant thyroid hormone receptor-beta Delta337T exclusively in cardiomyocytes [myosin heavy chain (MHC-mutant)]. These mice develop the cardiac hypothyroid phenotype in the presence of normal serum TH. Expression was also examined in wild-type mice rendered hypothyroid or hyperthyroid by pharmacological treatment. Atrial Cx40 mRNA and protein levels were decreased (85 and 55%, respectively; P < 0.001) in MHC-mt mice. Atrial and ventricular Cx43 mRNA levels were not significantly changed. Hypothyroid and hyperthyroid animals showed a 25% decrease and 40% increase, respectively, in Cx40 mRNA abundance. However, MHC-mt mice presented very low Cx40 mRNA expression regardless of whether they were made hypothyroid or hyperthyroid. Atrial depolarization velocity, as represented by P wave duration in electrocardiograms of unanesthetized mice, was extremely reduced in MHC-mt mice, and to a lesser extent also in hypothyroid mice (90 and 30% increase in P wave duration). In contrast, this measure was increased in hyperthyroid mice (19% decrease in P wave duration). Therefore, this study reveals for the first time that Cx40 mRNA is up-regulated by TH acting in cardiac atria via the TH receptor and that this may be one of the mechanisms contributing to atrial conduction alterations in thyroid dysfunctions.


A non-human primate model of acute liver failure suitable for testing liver support systems.
Acute hepatic failure is associated with high morbidity and mortality for which the only definitive therapy is liver transplantation. Some fraction of those who undergo emergency transplantation have been shown to recover native liver function when transplanted with an auxiliary hepatic graft that leaves part of the native liver intact. Thus, transplantation could have been averted with the development and use of some form of hepatic support. The costs of developing and testing liver support systems could be dramatically reduced by the availability of a reliable large animal model of hepatic failure with a large therapeutic window that allows the assessment of efficacy and timing of intervention. Non-lethal forms of hepatic injury were examined in combination with liver-directed radiation in non-human primates (NHPs) to develop a model of acute hepatic failure that mimics the human condition. Porcine hepatocyte transplantation was then tested as a potential therapy for acute hepatic failure. After liver-directed radiation therapy, delivery of a non-lethal hepatic ischemia-reperfusion injury reliably and rapidly generated liver failure providing conditions that can enable pre-clinical testing of liver support or replacement therapies. Unfortunately, in preliminary studies, low hepatocyte engraftment and over-immune suppression interfered with the ability to assess the efficacy of transplanted porcine hepatocytes in the model. A model of acute liver failure in NHPs was created that recapitulates the pathophysiology and pathology of the clinical condition, does so with reasonably predictable kinetics, and results in 100% mortality. The model allowed preliminary testing of xenogeneic hepatocyte transplantation as a potential therapy.


[Nutritional content of foods offered and consumed in a Spanish university canteen].
The prime objective of our work was to study the eating habits at lunchtime of staff and students at a University of "hidden due to confidentiality"of Spain. The second one was to attempt to reduce the energy consumption of cholesterol and fat in the diet of those groups. The study was made between 2010 and 2012 in the main canteen serving food at this university, focusing on food intake at lunch, the main meal of the day, containing between 35 and 40% of the total calories ingested throughout the day. A total of 9530 observations were made, each one corresponding to the nutritional valuation of food eaten (a complete lunch) per person, by students, teachers and service personnel. The study was carried out in 5 intervention stages and a previous non-intervention one to establish the habitual food intake of these groups. In each stage the nutritional information supplied to canteen users was increased to that in the final stage a modification of the price of the menus served was made. The food choices freely made by the groups were converted into the amount of nutrients by means of a prior nutritional evaluation of the dishes offered via their calculation using the database Nutriplato. This permitted the evaluation of 29 nutrients which, taking the user data observed, were converted into % of contribution to the RDA (per each population group), which produced a standardization of the data, permitting a subsequent statistical study by the GLM (SPSS v15) procedure to assess the effect of the different factors contemplated. The following were taken into account as classification factors (independent variables): sex, age group and level of body fat, as well as the intervention stage. The dependent variables were: energy, protein, lipid, carbohydrate, fiber, Ca, Mg, P, Na, K, Fe, Cu, Zn, Mn, I, Se, vitamins: B1, B2, B3, B5, B6, B8, B9, B12, C, A, D, E and cholesterol. The mean energy and cholesterol intake was significantly reduced in the individuals investigated in the last two intervention stages. Moreover, those last two stages were statistically differentiated, the last one being that which presented the lowest intake of those nutrients. As positive collateral effects, although this was not a priority objective in the study, a greater statistically significant intake of vitamin D, B9, dietetic fiber, and calcium and a lower intake of sodium with respect to the initial stages were noted. Our conclusions go in two directions: the positive effect of the nutritional information and the need to complement that information with economic measures that encourage still further food changes in population groups whose purchasing power is not very great.


Analysis of a yeast SNARE complex reveals remarkable similarity to the neuronal SNARE complex and a novel function for the C terminus of the SNAP-25 homolog, Sec9.
SNARE proteins represent a family of related proteins that are thought to have a central role in vesicle targeting and fusion in all eukaryotic cells. The binding properties of the neuronal proteins synaptobrevin 1 (VAMP1), syntaxin 1, SNAP-25, and soluble N-ethylmaleimide-sensitive factor attachment protein (alpha-SNAP), have been extensively studied. We report here the first biochemical characterization of a nonneuronal SNARE complex using recombinant forms of the yeast exocytic SNARE proteins Snc1, Sso1, and Sec9 and the yeast alpha-SNAP homolog, Sec17. Despite the low level of sequence identity, the association properties of the yeast and neuronal complexes are remarkably similar. The most striking difference we have found between the yeast and neuronal proteins is that individually neither of the target membrane SNAREs (t-SNAREs), Sso1 nor Sec9, show any detectable binding to the synaptobrevin homolog, Snc1. However, as a hetero-oligomeric complex, Sec9 and Sso1 show strong binding to Snc1. The clear dependence on the Sso1-Sec9 complex for t-SNARE function suggests that regulating the formation of this complex may be a key step in determining the site of vesicle fusion. In addition, we have used this in vitro assay to examine the biochemical effects of several mutations in Sec9 that result in pronounced growth defects in vivo. As expected, a temperature-sensitive mutation in the region most highly conserved between Sec9 and SNAP-25 is severely diminished in its ability to bind Sso1 and Snc1 in vitro. In contrast, a temperature-sensitive mutation near the C terminus of Sec9 shows no defect in SNARE binding in vitro. Similarly, a deletion of the C-terminal 17 residues, which is lethal in vivo, also binds Sso1 and Snc1 normally in vitro. Interestingly, we find that these same two C-terminal mutants, but not mutants that show SNARE assembly defects in vitro, act as potent dominant negative alleles when expressed behind a strong regulated promoter. Taken together these results suggest that the C-terminal domain of Sec9 is specifically required for a novel interaction that is required at a step following SNARE assembly.


The natural history of uveitis.
Inflammatory diseases of the eye were known to the ancients, but only recently have the underlying mechanisms to this problem become better defined. During the middle portion of this century, most cases of uveitis thought to be caused by infectious agents, such as those responsible for syphilis and tuberculosis. Since then, it has become clear that endogenous mechanisms of immunomodulation play an important role in these disorders, which along with environmental and genetic factors make up an important triad. Animals studies have indicated the pivotal role of the T-cell in many of these disorders. The development of T-cell lines has helped to further delineate cell to cell interactions that occur during an ocular inflammatory event. The presence in the eye of uveitogenic antigens raises the strong possibility of autoimmune driven processes as well, similar to what is seen in the animal models. The better understanding of ocular inflammatory mechanisms has led to improved therapeutic strategies, including Sandimmune, and more recently Cyclosporine G, a related compound that may be less nephrotoxic. Newer therapeutic strategies will focus on even more novel modes of immunomodulation, probably without the use of medications.


Sparse current source estimation for MEG using loose orientation constraints.
Spatially focal source estimates for magnetoencephalography (MEG) and electroencephalography (EEG) data can be obtained by imposing a minimum ℓ(1) -norm constraint on the distribution of the source currents. Anatomical information about the expected locations and orientations of the sources can be included in the source models. In particular, the sources can be assumed to be oriented perpendicular to the cortical surface. We introduce a minimum ℓ(1) -norm estimation source modeling approach with loose orientation constraints (ℓ(1) LOC), which integrates the estimation of the orientation, location, and strength of the source currents into a cost function to jointly model the residual error and the ℓ(1) -norm of the source estimates. Evaluation with simulated MEG data indicated that the ℓ(1) LOC method can provide low spatial dispersion, high localization accuracy, and high source detection rates. Application to somatosensory and auditory MEG data resulted in physiologically reasonable source distributions. The proposed ℓ(1) LOC method appears useful for incorporating anatomical information about the source orientations into sparse source estimation of MEG data.


Projected impact of vaccination timing and dose availability on the course of the 2014 west african ebola epidemic.
The 2014 West African Ebola outbreak has evolved into an epidemic of historical proportions and catastrophic scope. Prior outbreaks have been contained through the use of personal protective equipment, but such an approach has not been rapidly effective in the current epidemic. Several candidate vaccines have been developed against the Ebola virus, and are undergoing initial clinical trials. As removal of population-level susceptibility through vaccination could be a highly impactful control measure for this epidemic, we sought to estimate the number of vaccine doses and timing of vaccine administration required to reduce the epidemic size. Our base model was fit using the IDEA approach, a single equation model that has been successful to date in describing Ebola growth. We projected the future course of the Ebola epidemic using this model. Vaccination was assumed to reduce the effective reproductive number. We evaluated the potential impact of vaccination on epidemic trajectory under different assumptions around timing of vaccine availability. Using effective reproductive (Re) number estimates derived from this model, we estimate that 3-4 million doses of vaccine, if available and administered, could reduce Re to 0.9 in the interval from January-March 2015. Later vaccination would be associated with a progressively diminishing impact on final epidemic size; in particular, vaccination to the same Re at or after the epidemic is projected to peak (April-May 2015) would have little impact on final epidemic size, though more intensive campaigns (e.g., Re reduced to 0.5) could still be effective if initiated by summer 2015. In summary, there is a closing window of opportunity for the use of vaccine as a tool for Ebola epidemic control. Effective vaccination, used before the epidemic peaks, would be projected to prevent tens of thousands of deaths; this does not minimize the ethical challenges that would be associated with wide-scale application of vaccines that have undergone only limited evaluation for safety and efficacy.


Recent advances in mass spectrometry based clinical proteomics: applications to cancer research.
Cancer biomarkers have transformed current practices in the oncology clinic. Continued discovery and validation are crucial for improving early diagnosis, risk stratification, and monitoring patient response to treatment. Profiling of the tumour genome and transcriptome are now established tools for the discovery of novel biomarkers, but alterations in proteome expression are more likely to reflect changes in tumour pathophysiology. In the past, clinical diagnostics have strongly relied on antibody-based detection strategies, but these methods carry certain limitations. Mass spectrometry (MS) is a powerful method that enables increasingly comprehensive insights into changes of the proteome to advance personalized medicine. In this review, recent improvements in MS-based clinical proteomics are highlighted with a focus on oncology. We will provide a detailed overview of clinically relevant samples types, as well as, consideration for sample preparation methods, protein quantitation strategies, MS configurations, and data analysis pipelines currently available to researchers. Critical consideration of each step is necessary to address the pressing clinical questions that advance cancer patient diagnosis and prognosis. While the majority of studies focus on the discovery of clinically-relevant biomarkers, there is a growing demand for rigorous biomarker validation. These studies focus on high-throughput targeted MS assays and multi-centre studies with standardized protocols. Additionally, improvements in MS sensitivity are opening the door to new classes of tumour-specific proteoforms including post-translational modifications and variants originating from genomic aberrations. Overlaying proteomic data to complement genomic and transcriptomic datasets forges the growing field of proteogenomics, which shows great potential to improve our understanding of cancer biology. Overall, these advancements not only solidify MS-based clinical proteomics' integral position in cancer research, but also accelerate the shift towards becoming a regular component of routine analysis and clinical practice.


Standardized limb abduction for bioimpedance measurements using position restraints.
Bioelectrical impedance analysis (BIA) measurements are widely used in the assessment of body hydration. However, there are many variables which can influence the impedance values obtained and their individual significance is not entirely clear. One such variable is limb position and it is possible to standardize this either by invoking subject cooperation or by the use of some form of mechanical constraint. This study evaluates these two approaches. BIA measurements were made on five healthy male volunteers both with the positioners and without. Additional variables including room temperature, prandial status, exercise, alcohol intake, bed surface for the test, and bladder status were controlled. The mean percentage difference between impedance values with and without positioners was (-0.37 +/- 0.69)%. The mean difference between occasions with and without mechanical fixing was (2.02 +/- 2.5)% and (2.58 +/- 3)% respectively. None of these differences is statistically significant. It was concluded that reproducibility of BIA measurements obtained with active subject cooperation in limb position is not significantly improved if mechanical positioners are used. This may not apply where subject cooperation is poor e.g. in very sick or very young people or in repeated measurements in different days.


TRP64ARG beta 3-adrenergic receptor and obesity in Mexican Americans.
The beta 3-adrenergic receptor (beta 3AR) is expressed in visceral fat and is a regulator of resting metabolic rate, thermogenesis, and lipolysis. We genotyped 61 unrelated Mexican Americans for a variant in the beta 3AR gene (codon 64 TGGTrp-->CGGArg; TRP64ARG). The allele frequency was 0.13. The TRP64ARG variant was significantly associated with an earlier age of onset of non-insulin-dependent diabetes mellitus (41.3 +/- 4.6 years vs 55.6 +/- 2.6 years; P < 0.02) and in non-diabetics, with elevated 2-h insulin levels during an oral glucose tolerance test (810 +/- 120 pmol/l vs 384 +/- 6 pmol/l; P < 0.005). Non-diabetic subjects with the variant allele tended to have higher body mass indices (BMI), waist-to-hip ratios, and diastolic blood pressures. The study group was expanded to include 421 related subjects from 31 families in the San Antonio Family Diabetes Study. Using a measured genotype analysis approach to estimate genotype-specific means for each trait, those who were homozygous for the TRP64ARG variant had significantly higher 2-h insulin levels (P = 0.036) and trends towards higher BMI compared to the other two genotypes. We detected no associations of these traits in the TRP64ARG heterozygotes in the larger group. We conclude that the TRP64ARG beta 3AR variant is a susceptibility gene for several features of the insulin resistance syndrome in Mexican Americans. Since its effects are modest, study design (e.g., subject selection, genetic background, and statistical analyses) may influence which traits are associated with this variant and whether or not the effect is detectable in heterozygotes.


Generation of carbanions through stibine-metal and bismuthine-metal exchange reactions and its applications to precision synthesis of ω-end-functionalized polymers.
Generation of carbanions from organostibines and organobismuthines through heteroatom-metal exchange reactions was examined from synthetic and mechanistic viewpoints. The exchange reaction proceeded spontaneously upon treatment with various organometallic reagents, such as alkyl lithiums, tetraalkyl zincates, and alkyl magnesium halides to afford the corresponding carbanions quantitatively. Due to the high reactivity of these heteroatom compounds, the exchange reactions took place exclusively even in the presence of various polar functional groups, which potentially react with organometallic species. The advantage of this method was exemplified by the end-group transformation of living polymers that bear these heteroatom species at the ω-polymer end, prepared by using organostibine and bismuthine-mediated living radical polymerizations. Various polymers that bear polar functional groups and acidic hydrogen-for example, poly(methyl methacrylate), poly(butyl acrylate), poly(N-isopropyl acrylamide), and poly(2-hydroxyethyl methacrylate)-could be used in the exchange reactions, and subsequent trapping with electrophiles afforded the corresponding polymers with controlled molecular weights, molecular weight distributions, and end-group functionalities. Competition experiments showed that organostibines and organobismuthines were among the most reactive heteroatom compounds towards organometallic reagents and that their high reactivity was responsible for the high chemoselectivity in the exchange reaction.


Thiamin, riboflavin, and alpha-tocopherol content of exotic meats and loss due to gamma radiation.
Changes in thiamin, riboflavin, and alpha-tocopherol concentrations due to gamma irradiation were followed in alligator, caiman, bison, and ostrich (exotic) meats. The proximate composition showed that the exotic meats generally had lower fat content than domestic animal meats and that the thiamin content of the reptiles was lower. The changes in the vitamins due to irradiation were similar to those previously observed for domestic species. The results indicate that the loss of vitamins in these species is negligible insofar as the American diet is concerned, and that the concept of "chemiclearance" is applicable to exotic meats.


Segregated flux balance analysis constrained by population structure/function data: the case of PHA production by mixed microbial cultures.
In this study we developed a segregated flux balance analysis (FBA) method to calculate metabolic flux distributions of the individual populations present in a mixed microbial culture (MMC). Population specific flux data constraints were derived from the raw data typically obtained by the fluorescence in situ hybridization (FISH) and microautoradiography (MAR)-FISH techniques. This method was applied to study the metabolic heterogeneity of a MMC that produces polyhydroxyalkanoates (PHA) from fermented sugar cane molasses. Three populations were identified by FISH, namely Paracoccus sp., Thauera sp., and Azoarcus sp. The segregated FBA method predicts a flux distribution for each of the identified populations. The method is shown to predict with high accuracy the average PHA storage flux and the respective monomeric composition for 16 independent experiments. Moreover, flux predictions by segregated FBA were slightly better than those obtained by nonsegregated FBA, and also highly concordant with metabolic flux analysis (MFA) estimated fluxes. The segregated FBA method can be of high value to assess metabolic heterogeneity in MMC systems and to derive more efficient eco-engineering strategies. For the case of PHA-producing MMC considered in this work, it becomes apparent that the PHA average monomeric composition might be controlled not only by the volatile fatty acids (VFA) feeding profile but also by the population composition present in the MMC.