MCU overexpression evokes disparate dose-dependent effects on mito-ROS and spontaneous Ca2+ release in hypertrophic rat cardiomyocytes.
Cardiac dysfunction in heart failure (HF) and diabetic cardiomyopathy (DCM) is associated with aberrant intracellular Ca2+ handling and impaired mitochondrial function accompanied with reduced mitochondrial calcium concentration (mito-[Ca2+]). Pharmacological or genetic facilitation of mito-Ca2+ uptake was shown to restore Ca2+ transient amplitude in DCM and HF, improving contractility. However, recent reports suggest that pharmacological enhancement of mito-Ca2+ uptake can exacerbate ryanodine receptor-mediated spontaneous sarcoplasmic reticulum (SR) Ca2+ release in ventricular myocytes (VMs) from diseased animals, increasing propensity to stress-induced ventricular tachyarrhythmia. To test whether chronic recovery of mito-[Ca2+] restores systolic Ca2+ release without adverse effects in diastole, we overexpressed mitochondrial Ca2+ uniporter (MCU) in VMs from male rat hearts with hypertrophy induced by thoracic aortic banding (TAB). Measurement of mito-[Ca2+] using genetic probe mtRCamp1h revealed that mito-[Ca2+] in TAB VMs paced at 2 Hz under β-adrenergic stimulation is lower compared with shams. Adenoviral 2.5-fold MCU overexpression in TAB VMs fully restored mito-[Ca2+]. However, it failed to improve cytosolic Ca2+ handling and reduce proarrhythmic spontaneous Ca2+ waves. Furthermore, mitochondrial-targeted genetic probes MLS-HyPer7 and OMM-HyPer revealed a significant increase in emission of reactive oxygen species (ROS) in TAB VMs with 2.5-fold MCU overexpression. Conversely, 1.5-fold MCU overexpression in TABs, that led to partial restoration of mito-[Ca2+], reduced mitochondria-derived reactive oxygen species (mito-ROS) and spontaneous Ca2+ waves. Our findings emphasize the key role of elevated mito-ROS in disease-related proarrhythmic Ca2+ mishandling. These data establish nonlinear mito-[Ca2+]/mito-ROS relationship, whereby partial restoration of mito-[Ca2+] in diseased VMs is protective, whereas further enhancement of MCU-mediated Ca2+ uptake exacerbates damaging mito-ROS emission.NEW & NOTEWORTHY Defective intracellular Ca2+ homeostasis and aberrant mitochondrial function are common features in cardiac disease. Here, we directly compared potential benefits of mito-ROS scavenging and restoration of mito-Ca2+ uptake by overexpressing MCU in ventricular myocytes from hypertrophic rat hearts. Experiments using novel mito-ROS and Ca2+ biosensors demonstrated that mito-ROS scavenging rescued both cytosolic and mito-Ca2+ homeostasis, whereas moderate and high MCU overexpression demonstrated disparate effects on mito-ROS emission, with only a moderate increase in MCU being beneficial.


Method development to quantify Bv8 expression in circulating CD11b+ cells in patients with neovascular age-related macular degeneration (nvAMD) exhibiting Anti-VEGF refractoriness.
A subset of neovascular age-related macular degeneration (nvAMD) subjects appears to be refractory to the effects of anti-VEGF treatment and require frequent intravitreal injections. Prokineticin-2 (Bv8) expression in CD11b(+) cells has been linked to anti-VEGF response. We have developed a reproducible method to quantify gene expression in circulating CD11b + cells. Utilizing this method we tested the hypothesis that high Bv8 expression in circulating CD11b(+) cells is associated with anti-VEGF refractoriness in nvAMD patients. Two groups of nvAMD subjects undergoing treatment with anti-VEGF agents were recruited and classified as refractory or non-refractory to anti-VEGF treatment (n = 33 for each group). Two blood draws were obtained from each subject 1-9 months apart. Peripheral blood mononuclear cells (PBMCs) were isolated and CD11b(+) cells were purified via magnetic bead separation. RNA was purified, and relative expression of Bv8 among the subjects was compared via quantitative PCR analysis. Utilizing this approach no significant difference was detected in the mean LogRQ values between the first and second blood draws (t-test, p = 0.826) indicating low intra-patient variability and demonstrating good reproducibility of the assay. There was no significant difference in Bv8 expression between nvAMD subjects classified as refractory versus non-refractory. We were unable to find a correlation between Bv8 expression in CD11b + cells and anti-VEGF refractoriness in human nvAMD subjects. Relatively high expression in Bv8 in these subjects did not correlate with clinical treatment history, as measured by the frequency of injections. Utilizing this well characterized technique, studies are underway to examine alternative gene expression profiles in various circulating cell populations that may contribute to anti-VEGF refractoriness.


[Effect of smoking on the decline in forced expiratory volume in one second].
We used multiple regression analysis to study the effects of smoking cessation on the decline in forced expiratory volume in one second (FEV1). We studied 429 healthy men for about 3 years. Each subject was assigned to one of three groups, according to their self-reported smoking history: 281 were current smokers, 67 were former smokers, and 81 had never smoked. After statistical adjustment for initial age, initial FEV1, and decline in Maximum Mid-expiratory Flow (MMF), current smoking was found to be associated with a faster decline in FEV1. Having stopped smoking was associated with a slower decline in FEV1. The rate of decline in MMF was associated with the rate of decline in FEV1, which suggests that the rate of decline in FEV1 was greater in those subjects with small airways disease.


[Bladder tumor, metastasis from breast carcinoma. A case report with literature review].
Breast cancer is the most common type of cancer amongst. Norwegian women. It spreads to various localisations, the most usual foci being lymphatic nodes, lungs, liver and bone. For a long time, metastatic breast cancer to the urinary bladder has been considered an extremely rare manifestation. We describe one case in which metastasis to the bladder from breast cancer caused clinically important symptoms. We discuss the pathophysiology, symptoms and diagnostic tools in regard to this rare phenomenon, and why bladder metastasis from breast cancer has become, and will continue to become increasingly important.


Edelfosine and perifosine induce selective apoptosis in multiple myeloma by recruitment of death receptors and downstream signaling molecules into lipid rafts.
Multiple myeloma (MM) is an incurable B-cell malignancy, requiring new therapeutic strategies. We have found that synthetic alkyl-lysophospholipids (ALPs) edelfosine and perifosine induced apoptosis in MM cell lines and patient MM cells, whereas normal B and T lymphocytes were spared. ALPs induced recruitment of Fas/CD95 death receptor, Fas-associated death domain-containing protein, and procaspase-8 into lipid rafts, leading to the formation of the death-inducing signaling complex (DISC) and apoptosis. TNF-related apoptosis-inducing ligand receptor-1/death receptor 4 (TRAIL-R1/DR4) and TRAIL-R2/DR5, as well as Bid, were also recruited into lipid rafts, linking death receptor and mitochondrial signaling pathways. ALPs induced mitochondrial cytochrome c release. Bcl-X(L) overexpression prevented cytochrome c release and apoptosis. A Fas/CD95-deficient MM subline expressing DR4 and DR5 was resistant to edelfosine. Fas/CD95 retrovirus transduction bestowed edelfosine sensitivity in these cells. A Fas/CD95 mutant lacking part of the intracellular domain was ineffective. Lipid raft disruption prevented ALP-induced Fas/CD95 clustering, DISC formation, and apoptosis. ALP-induced apoptosis was Fas/CD95 ligand (FasL/CD95L) independent. ALP-induced recruitment of death receptors in lipid rafts potentiated MM cell killing by FasL/CD95L and TRAIL. These data uncover a novel lipid raft-mediated therapy in MM involving concentration of death receptors in membrane rafts, with Fas/CD95 playing a major role in ALP-mediated apoptosis.


Ultrafine Ru nanoparticles integrated on ordered mesoporous carbon for solvent-free hydrogenation of nitroarenes.
The hydrogenation of nitroarenes to aromatic amines with H2 under solvent-free conditions in place of organic solvents is a crucial process for the synthesis of fine chemicals. However, the catalysts that have been identified so far are relatively inactive with primary nitroarenes under solvent-free conditions. Herein, ordered mesoporous carbon (OMC)-supported highly dispersed Ru nanoparticles (Ru NPs) were easily prepared by using a coordination-assisted solvent evaporation induced co-assembly method, where 8-hydroxyquinoline exerts a significant influence on the mesostructure ordering and specific surface area of the Ru/OMC composites. The ultrasmall-sized Ru NPs (∼2.0 nm) supported on ordered mesoporous carbon were then applied in the hydrogenation of nitroarenes, exhibiting high activity and selectivity for numerous structurally diverse nitroarenes under solvent-free conditions. Compared with Ru/OMC-imp and Ru/AC-imp, Ru/OMC-800 exhibited a much higher activity and selectivity for hydrogenation of nitroarenes, suggesting its overwhelmingly better performance than OMC or AC supporting noble NPs by simple impregnation method. The strong metal-support interaction is capable of stabilizing the Ru NPs and achieving good recyclability as well as high selectivity.


Direct Measurement of the Angular Pair Correlation Coefficients in Molecular Liquids Using NMR. Benchmarking Force Fields for Atomistic Simulations.
High-field deuterium NMR spectroscopy is used to characterize a number of molecular liquids and their mixtures in order to probe the directional part of the intermolecular interactions through the orientational ordering induced in the isotropic liquid phase by the spectrometer magnetic field. The systems studied include benzene, chloroform, hexafluorobenzene, and thiophene at various concentrations and in mixtures. Dilution with the magnetically isotropic tetramethylsilane provides quantification of ordering at "infinite magnetic dilution", that is, in the absence of magnetic intermolecular correlations, and thereby allows identification of the contribution of these correlations to the orientational ordering in neat phases and at various degrees of magnetic dilution. Such contributions are conveyed by angular pair correlation coefficients, which, in addition to being accessible to direct NMR measurement, are also possible to evaluate directly from molecular dynamics simulations. By using various force fields, simulations provide benchmark quantities for testing and possibly further improving the force field performance, particularly with respect to the directional components of the intermolecular interactions. The latter are critical for the simulation of self-assembly generally and particularly in biological systems.


Evidence, illness, and causation: an epidemiological perspective on the Russo-Williamson Thesis.
According to the Russo-Williamson Thesis, causal claims in the health sciences need to be supported by both difference-making and mechanistic evidence. In this article, we attempt to determine whether Evidence-based Medicine (EBM) can be improved through the consideration of mechanistic evidence. We discuss the practical composition and function of each RWT evidence type and propose that exposure-outcome evidence (previously known as difference-making evidence) provides associations that can be explained through a hypothesis of causation, while mechanistic evidence provides finer-grained associations and knowledge of entities that ultimately explains a causal hypothesis. We suggest that mechanistic evidence holds untapped potential to add value to the assessment of evidence quality in EBM and propose initial recommendations for the integration of mechanistic and exposure-outcome evidence to improve EBM by robustly leveraging available evidence in support of good medical decisions.


THE EFFECT OF INVESTMENT IN ATTRACTIVE STRUCTURES ON ALLOCATION TO MALE AND FEMALE FUNCTIONS IN PLANTS.
Expressions for male and female fitnesses of partially self-fertilizing cosexual plants are derived, assuming that allocation to pollinator attraction at the time of flowering may decrease resources available for male and female primary structures. The total female fertility is assumed to be controlled by factors at two stages, flowering-time and fruiting-time, with resources for fruit maturation being limited so that maximum seed production may be limited by the availability of these resources. The fitness formulas are used to calculate ESS (evolutionarily stable strategy) allocations at flowering time to primary male and female sex functions and to attractive structures. These are compared with some data that are available for dry weights of different flower parts. The fitnesses of unisexual mutant forms are calculated, assuming that they are introduced into a population consisting mostly of the initial cosexual form and that they obey the same gain curves as that form. When compared with the fitness of the ESS cosexual form, this enables one to ask whether unisexual forms will be favored. We show that the spread of females is unlikely, unless there is high inbreeding depression and a rather high selfing rate, and that in some circumstances a linear relation between number of fertilized ovules and number of seeds matured can be less favorable for the invasion of females than is a highly concave relation. With a nearly linear relation between numbers of fertilized ovules and mature seeds, invasion by females is more likely when investment in attraction is low than when it is high. These effects are discussed in relation to the distribution of dioecy. The spread of male mutants is never likely in these models.


[COVID-19 Co-infection in a patient with Crimean Congo Hemorrhagic Fever: A Case Report].
Crimean-Congo Hemorrhagic Fever (CCHF) is an acute viral zoonotic disease. Coronavirus disease-2019 (COVID-19) is a newly emerging viral disease and it is caused by "severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2)". In this article, a case diagnosed with CCHF and COVID-19 coinfection confirmed by the polymerase chain reaction (PCR) method and its management was presented. A thirtyfive years old female patient admitted to the hospital with the complaint of fever for one day and common body pain. It was learned that three days before the onset of her complaints, she removed a tick adhering to the anterior abdominal wall with no precaution. Her body temperature was 38°C degrees and her respiratory rate was 22 per minute. The leucocyte count was 3660/mm³ and the platelet count was 138.000/mm³. It was determined that prothrombin time was 15.4 seconds, international normalized ratio (INR) was 1.35 seconds, and D-dimer level was 1310 ng/ml. The patient was hospitalized with prediagnosis of CCHF. Supportive treatment was started. On the second day at the clinical follow-up of the patient, complaints of sore throat and cough without sputum started. A combined nasopharyngeal and throat swab sample was taken from the patient because of the suspicion of COVID-19. COVID-19 PCR test result was reported as positive. Favipiravir treatment was started. The CCHF-PCR test, which was studied from the serum sample sent to the Microbiology Reference Laboratories was reported as positive. From the third day of favipiravir treatment; the patient did not have a fever and her complaints regressed. On the ninth day of her hospitalization, she was discharged. In this case; it is important to show that both diseases, especially in regions where CCHF disease is endemic, can be confused due to the similarity of the clinical picture with COVID-19 and to know that they can coexist.


The use of initial dosing of gentamicin in the management of pyelonephritis/urosepsis: A retrospective study.
The primary objective was to determine if initial empirical intravenous dosing of gentamicin improved patient's outcomes in pyelonephritis/urosepsis compared with alternative IV antibiotic management. Retrospective cross-sectional descriptive study. Public TertiaryTeaching Hospital serving adults in an urban centre. All adult patient records with a recorded diagnosis of any of pyelonephritis/urosepsis, urinary tract infection, UTI, complicated urinary tract infection, bacteriuria, symptomatic bacteriuria and asymptomatic bacteriuria from 2nd February 2012 to 10th May 2014 were reviewed. Only patients treated with an empirical regimen of one or more IV antibiotics were included in the study. The primary outcomes were: duration of IV antibiotic treatment, time to resolution of symptoms and length of hospital stay (LOS). Secondary end points were: compliance with Therapeutic Guidelines: Antibiotic (electronic version) (eTG) for severe pyelonephritis/urosepsis and appropriateness of gentamicin use. Univariate and multivariable associations between baseline and demographic variables and the main outcomes were performed using Chi-square tests and a General Linear Model using the SAS version 9.2 software. Of 295 patients reviewed 152 were prescribed one or more IV antibiotics and included in the study. Approximately half of the patients (n = 73, 48%) were prescribed IV piperacillin/tazobactam (Tazocin), while gentamicin was prescribed for 66 patients (43.4%). Of the 152 patients evaluated, 49 (32%) were given gentamicin first. Multivariable regression analysis showed that duration of IV treatment was shorter for those aged over 70 (40.2 hours vs 85.5 hours for those aged up to 70; p = 0.0074), and those who received gentamicin as first line treatment (41.3 hours vs 89.8 hours for those not receiving any gentamicin; p = 0.0312). After adjustment for age and gentamicin administration, there appeared to be no significant difference in duration of IV treatment for any other independent variables. No significant associations between the independent variables (gentamicin, age, gender, comorbidities, and eTG compliance) and either time to resolution of symptoms (median: 68 hours) or hospital LOS (median: 5 days) were observed. Neither time to resolution of abnormal symptoms nor length of stay were significantly influenced by an initial dose of gentamicin when the data were subjected to multivariable analysis. The age of the patient and pattern of gentamicin treatment were the dominant factors associated with duration of IV antibiotics. Piperacillin/tazobactam is not recommended in treatment guidelines for pyelonephritis/urosepsis but was the most commonly prescribed IV antibiotic. This requires review by the appropriate hospital clinicians.


Reconstructing multivariate causal structure between functional brain networks through a Laguerre-Volterra based Granger causality approach.
Classical multivariate approaches based on Granger causality (GC) which estimate functional connectivity in the brain are almost exclusively based on autoregressive models. Nevertheless, information available from past samples is limited due to both signal autocorrelation and necessarily low model orders. Consequently, multiple time-scales interactions are usually unaccounted for. To overcome these limitations, in this study we propose the use of discrete-time orthogonal Laguerre basis functions within a Wiener-Volterra decomposition of the BOLD signals to perform effective GC assessments of brain functional connectivity. We validate our method in synthetic noisy oscillator networks, and analyze experimental fMRI data from 30 healthy subjects publicly available within the Human Connectome Project (HCP). Synthetic results demonstrate that our Laguerre-Volterra based GC estimates outperform classical approaches in terms of accuracy in detecting true causal links while rejecting false causal links in complex nonlinear networks. Human data analysis shows for the first time that the default mode network modulates both the salience network as well as fronto-temporal circuits in a causal fashion.


Trajectories of symptom dimensions in short-term response to antipsychotic treatment in patients with a first episode of non-affective psychosis.
Trajectory patterns of positive, disorganized and negative dimension symptoms during antipsychotic treatment in drug-naive patients with first-episode psychosis have yet to be examined by using naturalistic data. This pragmatic clinical trial randomized 161 drug-naive patients with a first episode of psychosis to olanzapine, risperidone or haloperidol. Patients were assessed with the Scale for the Assessment of Negative Symptoms (SANS) and Positive Symptoms (SAPS) at baseline and at the end of weeks 1, 2, 3, 4 and 6 of antipsychotic treatment. Censored normal models of response trajectories were developed with three dimensions of the SAPS-SANS scores (positive, disorganized and negative) in order to identify the different response trajectories. Diagnosis, cannabis use, duration of untreated psychosis (DUP), smoking and antipsychotic class were examined as possible predictive variables. Patients were classified in five groups according to the positive dimension, three groups according to the disorganized dimension and five groups according to the negative dimension. Longer DUPs and cannabis use were associated with higher scores and poorer responses in the positive dimension. Cannabis use was associated with higher scores and poorer responses in the disorganized dimension. Only schizophrenia diagnosis was associated with higher scores and poorer responses in the negative dimension. Our results illustrate the heterogeneity of short-term response to antipsychotics in patients with a first episode of psychosis and highlight markedly different patterns of response in the positive, disorganized and negative dimensions. DUP, cannabis use and diagnosis appeared to have a prognostic value in predicting treatment response with different implications for each dimension.


An Enantioselective Total Synthesis of (+)-Duocarmycin SA.
An efficient, concise enantioselective total synthesis of the potent antitumor antibiotic (+)-duocarmycin SA is described. The invented route is based on a disconnection strategy that was devised to facilitate rapid and efficient synthesis of key core compounds to enable preclinical structure-activity relationship investigations. The key tricycle core was constructed with a highly enantioselective indole hydrogenation to set the stereocenter and a subsequent hitherto unexplored vicarious, nucleophilic-substitution/cyclization sequence to effectively forge a final indole ring. Additionally, the development of a stable sulfonamide protecting group capable of mild chemoselective cleavage greatly enhanced sequence yield and throughput. An understanding of key reaction parameters ensured a robust, reproducible sequence easily executable on decagram scales to this highly promising class of compounds.


Peroxiredoxin 6 is upregulated in bovine oocytes and cumulus cells during in vitro maturation: role of intercellular communication.
Peroxiredoxins are peroxidases involved in antioxidant defense and intracellular signaling. Expression of transcripts coding for peroxiredoxin 6 (PRDX6) has been previously described to be upregulated in oocytes after in vitro maturation, a period during which general transcription decreases dramatically in oocytes. The aim of the present work was to evaluate PRDX6 regulation in bovine cumulus-oocyte complexes in relation to maturation and intercellular communication. PRDX6 expression was analyzed by reverse transcription-PCR and Western blotting in oocytes and cumulus cells before and after in vitro maturation. PRDX6 was found to be upregulated at the mRNA and protein levels in both cell types after maturation. The effect of paracrine and gap junctional communication on PRDX6 expression was then assessed by culturing cumulus clusters in the presence or absence of denuded oocytes. While PRDX6 upregulation in oocytes required intact cumulus-oocyte junctions, the presence of denuded oocytes was necessary but sufficient for the upregulation to occur in cumulus cells. Finally, the influence of recombinant mouse growth differentiation factor-9 (GDF-9) on PRDX6 expression in cumulus cells was studied. GDF-9 induced cumulus expansion and PRDX6 upregulation in bovine cumulus clusters. Altogether, our data suggest that PRDX6 upregulation in cumulus-oocyte complexes during in vitro maturation is mutually regulated by both cell types: PRDX6 upregulation in oocytes would require gap junctions with cumulus cells, while upregulation in cumulus would depend on secretion of oocyte paracrine factor(s) with GDF-9 being a likely candidate.


Comparison of orthostatic reactions of patients still unconscious within the first three months of brain injury on a tilt table with and without integrated stepping. A prospective, randomized crossover pilot trial.
To determine whether passive leg movement during tilt table mobilization reduces the incidence of orthostatic dysfunction in mobilization of patients being comatose or semi-comatose early after brain injury. Randomized crossover pilot trial using sequential testing. Neurorehabilitation hospital. Nine patients still unconscious within the first three months of brain injury (5 men, 4 women; age 51 +/- 20 years). Patients were subjected once to a conventional tilt table and once to a tilt table with an integrated stepping device. The number of syncopes/presyncopes (orthostatic hypotension, tachypnoea, increased sweating) during interventions. One patient had presyncopes on both devices, six patients had presyncopes on the conventional tilt table but not on the tilt table with integrated stepping, and two patients did not exhibit presyncopal symptoms on either device. There were significantly more incidents on the tilt table without than on the one with an integrated stepping device (P < 0.05) at tilts of 50 or 70 degrees respectively. Patients tolerate greater degrees of head-up tilt better with simultaneous leg movement.


Double umbilical cord blood transplant is effective therapy for relapsed or refractory Hodgkin lymphoma.
A sub-group of patients with Hodgkin Lymphoma (HL) who relapse after autologous stem cell transplant can achieve long-term disease-free-survival after allogeneic stem cell transplant (alloSCT). There is limited information regarding the tolerability and efficacy of double umbilical cord blood transplant (dUCBT) for relapsed/refractory HL. We analyzed 27 consecutive, heavily pre-treated patients receiving dUCBT for relapsed/refractory HL at two centers from 2003-2014. The majority of patients relapsed <6 months after autologous stem cell transplant. A total of 15 patients received myeloablative (most commonly melphalan, fludarabine, thiotepa and anti-thymocyte globulin [ATG]) and 12 non-myeloablative conditioning regimens (fludarabine, cyclophosphamide, 200cGy total body irradiation +/- ATG). All patients engrafted; median time to neutrophil and platelet engraftment was 17 and 37 days, respectively. Overall response rate was 68%; 58% achieved complete remission. Median progression-free survival (PFS) was 12.2 months; median overall survival was 27 months. Cumulative incidences of relapse and of non-relapse mortality at 5 years were 30% and 37.9%, respectively; 5-year PFS was 31.3% (95%CI 10.1-52.5). There was a trend toward inferior PFS in patients with lymph node size ≥2 cm at the time of alloSCT (p = 0.07) and toward inferior survival in patients with chemorefractory disease pre-alloSCT (p = 0.12). dUCBT is feasible in patients with heavily pre-treated HL and can achieve long-term disease-free survival in approximately 30% of patients.


The Proportion of ALDEFLUOR-Positive Cancer Stem Cells Changes with Cell Culture Density Due to the Expression of Different ALDH Isoforms.
A significant number of discrepancies exist within the literature regarding ALDEFLUOR-positive stem cell populations in cell lines. We hypothesized that these inconsistencies resulted from differences in culture conditions, particularly cell density. We cultured several colon cancer cell lines (N=8) at high and low densities and found a significant decrease in ALDEFLUOR-positive cell populations at high density. However, we found no changes in the CD166-positive stem cell population, self-renewal, or cell cycle distribution of cells cultured at different densities. Interestingly, when we sorted both ALDEFLUOR positive and negative populations from the different density cultures, we identified a significant number of Aldehyde dehydrogenase (ALDH) isoforms whose expression was decreased in ALDEFLUOR-positive stem cells cultured at high density. This novel finding suggests that multiple ALDH isoforms contribute to ALDEFLUOR activity in colon cancer stem cells and decreases in ALDEFLUOR-positive stem cells at high cell density are due to decreased expression of multiple ALDH isoforms. Thus, designing therapeutics to target ALDEFLUOR-positive cancer stem cells may require inhibition of multiple ALDH isoforms.


[Critical analysis of the immunological self/non-self model and of its implicit metaphysical foundations].
An examination of the concepts used in immunology prompts us to wonder about the origins and the legitimacy of the notions of self and non-self, which constitute the core of the dominant theoretical model in this science. All theoretical reflection concerning immunology must aim at determining a criterion of immunogenicity, that is, an operational definition of the conditions in which an immune reaction occurs or does not occur. By criticizing both conceptually and experimentally the self/non-self vocabulary, we can demonstrate the inaccuracy and even the inadequacy of the dichotomy of self/non-self. Accordingly, the self/non-self model must be reexamined, or even rejected. On the basis of this critique, we can suggest an alternative theoretical hypothesis for immunology, based on the notion of continuity. The 'continuity hypothesis' developed here attempts to give a criterion of immunogenicity that avoids the reproaches leveled at the self model.


CHI study: protocol for an observational cohort study on ageing and mental health in community-dwelling older adults.
Ageing is associated with a multitude of healthcare issues including dementia, depression, frailty, morbidity associated with chronic disease and high healthcare utilisation. With Singapore's population projected to age significantly over the next two decades, it has become increasingly important to understand the disease burden and etiological process among older adults. The Community Health and Intergenerational study aims to holistically examine ageing in place by investigating the resilience and vulnerability factors of the ageing process in the biological, psychological and social domains within the environment. Using a cohort multiple randomised controlled trial design, comprehensive health profiles of community-dwelling older adults will be collected. The objective is to recruit 1000 participants (aged 60-99 years) living in the western region of Singapore within a period of 3 years (2018-2020). Assessments include basic sociodemographic, physical health and function (cardiac, oral and blood profiles and visual function), cognitive functioning, daily functioning, physical fitness, emotional state, free-flowing speech, sleep quality, social connectedness, caregiver burden, intergenerational communication, quality of life, life satisfaction, attitudes to ageing and gratitude and compassion. Results from the cohort will enable future studies to identify at-risk groups and develop interventions to improve the physical and mental health and quality of life of older adults. Approval of the cohort study by the National University of Singapore Institutional Review Board (NUS-IRB Reference code: H-17-047) was obtained on 12 October 2017. Written consent will be obtained from all participants. Findings from the cohort study will be disseminated by publication of peer-reviewed manuscripts, presentations at scientific meetings and conferences with local stakeholders.


[Migration of a Temporary Epicardial Pacing Lead into the Vascular Graft].
A 48-year-old man was refered to our hospital for the treatment of bicusp aortic valve and severe aortic regurgitation. He underwent Bentall procedure (SJM 25mm+HemashieldØ 28 mm composite graft). Temporary epicardial pacing lead( TPL) was placed on the right ventricle at the opetation, and was cut on his skin surface prior to discharge because of difficulty in traction removal. After 15 years from operation, follow up computered tomography and echocardiography showed migration of 2 retaining TPLs extending from vascular graft of ascending aorta to the right innominate artery. We performed Redo-median sternotomy and removed the TPLs. This is the 1st reported case of a migrated temporary pacing lead into the vascular graft of ascending aorta under noninfectious conditions.


Combination of magnetic and chemocytotoxic cancer cell depletion for autologous bone marrow transplantation.
An artificial mixture of breast cancer cell line cells (T-47D) and normal human marrow cells was used to investigate optimal approaches for autologous BMT. The experiments were designed to test the applicability of chemocytotoxic agents, a dye-mediated photolytic agent and SBA for in vitro purging in autologous BMT of patients with advanced malignancies. Treatment with high concentration of etoposide (VP-16) (10 to 80 micrograms/ml) resulted in a maximal depletion of 1.5 log, whereas more efficient tumor cell eradication (2.5 log) was achieved by 30-min incubation with 100 micrograms/ml4HC. Photosensitization by exposure for 90 min to daylight in the presence of MC-540 could remove only 1 log of T-47D cells. The chemocytotoxic treatment with 4HC was chosen to follow initial tumor cell separation by SBA bound to polystyrene magnetic beads that had previously been shown to bind to several cancer cell types while sparing marrow progenitor cells. Artificial mixtures containing 10 to 14% T-47D cells in fresh normal BM cells were subjected to SBA-magnetic beads, and the SBA-negative fraction was further treated with 4HC. The combined two-step procedure resulted in a consistent tumor cell depletion of greater than 4 logs. The purging procedure appears acceptable for clinical marrow purging prior to cyropreservation and autotransplantation in patients with documented BM involvement of neoplastic cells sensitive to 4HC with positive binding to SBA such as breast cancer.


[Childhood trauma and its correlation with resilience among primary and middle school students in Wuhan city in 2015].
To investigate the prevalence of early trauma and resilience among adolescents in Wuhan, and explore the relationship between early trauma and resilience. Totally 4871 students aged 10-16 years were chosen by cluster sampling in Wuhan city from September to October 2015. All subjects completed self-report questionnaires, including general information, the Childhood Trauma Questionnaire( CTQ), and the Connor Davidson Resilience Scale( CD-RISC). The mean score of CD-RISC of the total sample was( 64. 70 ± 18. 34). Statistical significance in different gender( t = 5. 373, P<0. 001), age( F = 49. 401, P<0. 001), single child( t = 3. 529, P<0. 001), levels of mother's education( F = 36. 129, P< 0. 001), relationship between parents( F = 89. 831, P < 0. 001), family economic status( F = 36. 547, P<0. 001). The rate of early trauma was 30. 1%. Male( χ~2= 42. 272, P < 0. 001), lower levels of mother 's education( χ~2= 44. 345, P < 0. 001), poorer relationship between parents( χ~2= 133. 045, P < 0. 001), and worse family economic status( χ~2= 31. 231, P<0. 001) were associated with increased risk of early trauma. The scores of emotional abuse, physical abuse, sexual abuse, emotional neglect and physical neglect were negatively correlated with the scores of CD-RISC( r followed by-0. 256, -0. 107, -0. 053, -0. 355 and-0. 308, P<0. 01). Regression analysis implied female, older age, emotional abuse, emotional neglect, and physical neglect( B followed by-0. 156, -0. 117, -0. 109, -0. 214 and-0. 149, P < 0. 01) of primary and middle school students assumed predictive resilience. Emotional abuse, emotional neglect, and physical neglect are negatively associated with resilience among children and adolescent. The result suggest that reducing emotional abuse, emotional neglect and physical neglect experience may contribute to child resilience.


Antidepressants in adult suicides in New York City: 2001-2004.
The U.S. Food and Drug Administration recently extended the Black Box warning on anti-depressants regarding pediatric suicidality to include young adults. The decision was guided by results from meta-analyses of 372 randomized controlled clinical trials of antidepressants for adults. Nearly all suicidality in those trials was nonfatal suicide attempts and ideation. Here, we consider whether antidepressants are linked with adult suicide deaths. Subjects in this medical examiner surveillance study included all suicides, 18 years and older, in New York City from 2001-2004. Postmortem blood was analyzed for the presence of antidepressants. There were 1419 adult suicides in New York City during the study period. Antidepressants were detected at autopsy in 23.1% of the suicides who met criteria for toxicology analyses. Antidepressants were least prevalent in suicides aged 18-24 years (13.9%). Antidepressants were detected in less than one-quarter of adult suicides in New York City from 2001-2004. The majority of the suicides were not attributable to antidepressant use, and perhaps many could have been prevented with appropriate treatment. Although this study does not provide evidence for a link between antidepressant use and subsequent suicide, careful monitoring of patients receiving antidepressants remains critically important.


Suppression of LH response to gonadotrophin-releasing hormone or oestradiol by ACTH(1-24) treatment in anoestrous ewes.
Stress is known to result in lowered female reproductive efficiency. The objective of this study was to examine how increased pituitary-adrenal activity may influence gonadotrophin release in anoestrous ewes. Various doses (0.06-1.0 mg) of a synthetic adrenocorticotrophic hormone (ACTH(1-24)) preparation were injected into ewes 30 min or 3 h before an i.v. injection of 500 ng gonadotrophin-releasing hormone (GnRH). The LH response to GnRH given 30 min after ACTH(1-24) was similar to that after GnRH alone, whereas the response 3 h after ACTH(1-24) was significantly lower, irrespective of the dose of ACTH(1-24). At 30 min and 3 h after ACTH(1-24) the concentrations of cortisol exceeded 50 nmol/l compared with baseline values of less than 10 nmol/l. The effect of ACTH(1-24) on oestradiol-induced LH release was also examined. Those ewes receiving 0.8 mg ACTH(1-24) depot and 50 micrograms oestradiol benzoate simultaneously had a preovulatory-type increase in LH 14-20 h later, similar to when oestradiol benzoate was given alone. None of the ewes receiving an additional 0.8 mg ACTH(1-24) depot 10 h after oestradiol benzoate had increases in LH concentration. The cortisol concentrations in all ewes receiving either one or two injections of ACTH(1-24) were greater than 35 nmol/l at 10 h after the oestradiol injection. However, concentrations of progesterone increased from 0.9 +/- 0.3 (S.E.M.) nmol/l at the time of the second ACTH(1-24) injection to 2.1 +/- 0.3 nmol/l after 2 h.(ABSTRACT TRUNCATED AT 250 WORDS)


Investigations of the effect of nonmagnetic Ca substitution for magnetic Dy on spin-freezing in Dy₂Ti₂O₇.
Physical properties of partially Ca substituted hole-doped Dy2Ti2O7 have been investigated by ac magnetic susceptibility χ(ac)(T), dc magnetic susceptibility χ(T), isothermal magnetization M(H) and heat capacity C(p)(T) measurements on Dy1.8Ca0.2Ti2O7. The spin-ice system Dy2Ti2O7 exhibits a spin-glass type freezing behavior near 16 K. Our frequency dependent χ(ac)(T) data of Dy1.8Ca0.2Ti2O7 show that the spin-freezing behavior is significantly influenced by Ca substitution. The effect of partial nonmagnetic Ca(2+) substitution for magnetic Dy(3+) is similar to the previous study on nonmagnetic isovalent Y(3+) substituted Dy(2-x)Y(x) Ti2O7 (for low levels of dilution), however the suppression of spin-freezing behavior is substantially stronger for Ca than Y. The Cole-Cole plot analysis reveals semicircular character and a single relaxation mode in Dy1.8Ca0.2Ti2O7 as for Dy2Ti2O7. No noticeable change in the insulating behavior of Dy2Ti2O7 results from the holes produced by 10% Ca(2+) substitution for Dy(3+) ions.


Molecular identification of the Marek's disease virus vaccine strain CVI988 in vaccinated chickens.
The study describes three polymerase chain reaction (PCR) systems for the CVI988 vaccine virus: the meq gene, the MDV BamHI-D/H 132 bp tandem repeat fragment and the MDV-gB gene. Whereas the PCR product of virulent MDV strains and of the CVI988 virus strain with the meq and the 132 bp primer sets differed for the two templates, the MDV-gB PCR products were similar. The sensitivity of the three PCRs was determined for the two templates: the CVI988 DNA was detected up to 2.48 plaque forming units, and a MDV-1 DNA, was amplified with the 132 bp primers up to the 10(-3) DNA dilution, and up to the 10(-2) with the MDV-gB and meq gene primers. As conventional detection for the CVI988 vaccine virus is by tissue culture, the aim was to analyse the feasibility of the molecular detection of the vaccine virus in the vaccinated chick. In two experimental trials employing specific pathogen free and commercial Lohmann chicks, respectively, the vaccine virus replicated to a limited extent; it was detected only in the spleen of up to 60% chicks at 2-4 weeks and in one chick at 3 weeks, respectively. The survey of three commercial Lohmann flocks, kept in biosecurity conditions, revealed the vaccine virus only in the spleen of 40% of 30-day-old chicks. The present study shows that CV1988 DNA is present in vaccinated chicks in a low quantity and it is difficult to detect directly from the chick, probably because vaccine viruses are latent in vivo. For an efficient detection it is pertinent to cultivate the vaccine virus on chicken embryo fibroblasts (CEF), as then the virus escapes the latent state, enters into the productive mode of replication, and a high viral copy number is produced.


Susceptibility of anaerobic bacteria to FCE 22101.
The activity of FCE 22101 against 675 strains of anaerobic bacteria was determined by an agar dilution method. Its activity was compared with those of benzylpenicillin, piperacillin, cefoxitin, imipenem, clindamycin, metronidazole, chloramphenicol, vancomycin, fusidic acid, and bacitracin. FCE 22101 and imipenem (MIC, less than or equal to 0.5 mg/liter) were the most active agents against the anaerobic strains tested, except against Clostridium difficile strains. On the basis of these results, FCE 22101 appears to be a promising antimicrobial agent for treatment of anaerobic infections, and further clinical investigations are indicated.


A multielement trace mineral injection improves liver copper and selenium concentrations and manganese superoxide dismutase activity in beef steers.
Trace minerals (TM) are vital to health and growth of livestock, but low dietary concentrations and dietary antagonists may reduce mineral status and feeder cattle TM status is usually unknown at arrival. The objective of this study was to examine the effect of TM status on response to mineral injection in beef cattle. Forty steers were equally assigned to diets for an 84-d depletion period: control (CON; supplemental Cu, Mn, Se, and Zn) or deficient (DEF; no supplemental Cu, Mn, Se, or Zn plus Fe and Mo as TM antagonists). Lesser liver Cu and Se concentrations (79.0 ± 11.60 and 1.66 ± 0.080 mg/kg DM, respectively) in DEF steers compared with CON steers (228.8 ± 11.60 and 2.41 ± 0.080 mg/kg DM, respectively) on d 71 of depletion indicated mild deficiencies of these TM (P < 0.001). On d 1 of the 85-d repletion period, 10 steers within each dietary treatment were injected with sterilized saline (SAL) or Multimin90 (MM), containing 15, 10, 5, and 60 mg/mL of Cu, Mn, Se, and Zn, respectively, at a dose of 1 mL/68 kg BW. All steers were fed the same repletion diet supplemented with Cu, Mn, Se, and Zn to meet or exceed NRC recommendations. Blood was collected on d 0 and 1, and blood and liver biopsies were collected on d 8, 15, 29, 57, and 85 postinjection. Red blood cell lysate manganese-superoxide dismutase activity was greater in MM (P = 0.02), suggesting incorporation of injectable TM into a biological process. The increase in liver Se in response to MM was greater in CON vs. DEF (P = 0.02), suggesting TM from injection were used rather than stored in DEF steers. Liver Se and Cu (P < 0.05) were elevated through at least d 30 by MM. Dietary TM deficiency decreased neutrophil bacteria killing ability and increased myeloperoxidase (MPO) degranulation (P < 0.04) as measured on d 0, 1, 13, and 14 during the repletion period while injection had no impact. Within CON animals, total MPO was greater in animals that received TM injection, but injection did not affect MPO within DEF steers (P = 0.007). Overall, TM from an injectable mineral were used differently between TM adequate and mildly deficient steers.


Changing picture of leprosy in North Arcot District, Tamil Nadu after M.D.T.
This paper discussed the effect in a 5 year period (1983-88) of MDT on the Leprosy situation in North Arcot District where MDT was started in 1983. The cases at the start of MDT were 68351 and 29511 cases were detected in the year period and the total case load was 97862. Out of this total case load 84810 cases were deleted by RFC, deaths or PL*. The causes for deletions are discussed in detail. The remaining case load at the end of the 5 year period is 13052 or 13.35% of the cases at start. The M.D.T. has definitely played a part in the drastic reduction of the case load since the major number of cases have been deleted as RFC due to cure of the disease and so the future planning of leprosy work when the case load becomes very low is also discussed.


The effects of repeated antenatal glucocorticoid therapy on the developing brain.
Antenatal glucocorticoid (GC) therapy improves infant outcome following preterm birth. As approximately 50% of women given a first course of antenatal GCs remain undelivered 7-14 d later, many clinicians administer further courses. GCs are known to be neurotoxic and there is concern that exposure during early development may have adverse effects on the immature brain. The aim of this investigation was to compare magnetic resonance (MR) indices of brain maturation in infants exposed to repeated antenatal GC therapy and born at or close to term, with non-GC exposed control infants. MR images were obtained during quiet sleep without sedation. T1 weighted volume images were obtained in the sagittal plane and T1, T2 weighted spin echo and inversion recovery images in the transverse plane. Brain volume and surface area were calculated from segmented image slices, and a measure of the complexity of cortical folding, the whole cortex convolution index (WCCI), from computerized analysis of a vector coded contour following algorithm. Analysis of covariance was used to compare the two groups after allowing for the effect of postmenstrual age. There were 10 infants in the GC group (range of antenatal GC exposure, 3-11 courses) and 6 controls. Each GC course comprised two 12-mg IM doses of betamethasone 24 h apart. GC exposed infants had a significantly lower WCCI (p = 0.001) and smaller surface area (p = 0.02), after allowing for postmenstrual age. There was no significant difference in brain volume (p = 0.5). Repeated antenatal GC exposure results in measurable differences in brain maturation when compared with gestational age matched non-GC exposed controls. The clinical relevance of these observations is not known.


A Microplate-Based Bioluminescence Assay of Mitochondrial Calcium Uptake.
Mitochondrial Ca2+ homeostasis is crucial for regulating vital functions such as respiration or apoptosis. Targeted aequorins are excellent probes to measure subcellular Ca2+. Ca2+ concentration in mitochondria ([Ca2+]M) is low at rest (about 10-7 M) and can increase to the micromolar or even approach the millimolar range, upon cell activation. Here we describe a new quantitative luminescent protocol to directly measure mitochondrial Ca2+ uptake, optimized for high throughput. The sensitivity of the method allows detection of changes in either the capacity or the affinity of mitochondrial Ca2+ transport.


Medical Admissions Among Adolescents With Eating Disorders During the COVID-19 Pandemic.
Emerging data suggest the coronavirus disease 2019 (COVID-19) pandemic has been associated with worsening symptoms of eating disorders (EDs) among both adults and adolescents. With this study, we sought to determine if medical admission patterns among adolescents admitted to our institution for restrictive EDs changed during the pandemic, relative to prepandemic counts of admissions per month. We performed a chart review of patients aged 10 to 23 years admitted to our children's hospital for restrictive EDs from March 2017 through March 2021 and completed an interrupted time series analysis of admission counts per month. Demographic variables for admitted patients were compared by using χ2, Fisher's exact, and 2-sample t tests. ED-related medical admissions at our institution increased significantly during the COVID-19 pandemic. The total number of admissions during the first 12 months of the COVID-19 pandemic (April 1, 2020, through March 31, 2021, n = 125) was more than double the mean number of admissions per year for the same time frame (April 1 through March 31) for the previous 3 years (mean = 56). Patient demographics were similar before and during the pandemic, with the exception that patients admitted during the COVID-19 pandemic were less likely than those admitted before the pandemic to have public insurance. Medical admissions related to restrictive EDs among adolescents increased significantly during the COVID-19 pandemic. Pediatric providers in a variety of settings should be prepared to care for adolescents with restrictive EDs during the pandemic.


Cardiovascular disease risk factors and attitudes towards prevention among Korean-American elders.
The leading cause of death among the aging population is cardiovascular disease. Cardiovascular disease prevention and modification of disease risk factors are important and worthwhile directions for study because this population is rapidly increasing. Past studies have shown the significance of modifying cardiovascular disease risk factors in Anglo populations where major risk factors of hypertension, elevated cholesterol levels, diabetes, obesity, physical inactivity and smoking have been identified. However, the prevalence of these risk factors and the attitudes towards modifying them have not been studied in the Korean-American elderly population. This preliminary study used questionnaires given to fifty older recently immigrated Korean-American participants of a Korean senior center. Results show that this population has a lower frequency of cardiovascular disease and all risk factors, except for diabetes compared to an Anglo elderly population In addition, older Korean-American elders were generally unaware of risk factors for cardiovascular disease and uninterested in changing their habits to avoid cardiovascular disease. Increased availability of educational campaigns and health care in Korean are recommended.


Salmonella in Victoria, 1997: the story so far.
The Infectious Diseases Unit of the Department of Human Services, Victoria, reported an increased incidence of Salmonella infections in early 1997. To 21 April 1997, 944 notifications had been received, passing the previous year's total of 915. Five outbreaks of five separate serovars have been investigated and traced to their sources. The outbreaks, their sources and the control measures undertaken are described. Further clusters of other Salmonella serovars are being investigated.


Working with older people from minority ethnic groups.
Living in a multi-cultural community means nurses in all specialties must have an understanding of the diverse ethnic backgrounds of their patients. This applies equally to nurses working with older people, who can enhance the care they give by facilitating adequate, appropriate and accessible services for all patients, irrespective of ethnic origin.


Development and validation of the Clinical Aspects of Historical Trauma Questionnaire in Rwandan genocide survivors.
Historical trauma is a relatively new concept in the literature, and investigations are needed to clarify its clinical aspects and develop instruments to measure its sequelae. The purpose of this study was to develop the Clinical Aspects of Historical Trauma Questionnaire (CAHTQ), which is meant to capture trauma sequelae in different contexts, and provide initial psychometric information. Participants were survivors of the genocide against the Tutsi in Rwanda (N = 261) aged 32-87 years (M = 46.30 years, SD = 11.95) who completed a preliminary version of the CAHTQ, constructed based on theoretical and content-related consideration, as well as the International Trauma Questionnaire, Fatalism Scale, Public Health Depression Questionnaire, Brief Coping Inventory, Forgiveness Questionnaire, and Sentiment of Reconciliation Questionnaire to test the discriminant and convergent validity of the CAHTQ. Exploratory factor analysis was conducted to reduce the number of items and extract factors; confirmatory factor analysis (CFA) was conducted to confirm the measure's dimensionality. The final questionnaire includes 20 items and five subscales. The items demonstrated good internal consistency, Cronbach's α = .91, and the CFA demonstrated a very good fit of the model to the data, χ2 (60, N = 261) = 271, CFI = .963 = , TLI = .956, SMRR = .052, RMSEA = .052. The CAHTQ was developed to capture the clinical aspects of historical trauma sequelae. Unlike comparable previously developed instruments, this questionnaire can be used for various historical traumas globally, and its suitability for this purpose will be the focus of future studies.


Dansylglycine, a fluorescent probe for specific determination of halogenating activity of myeloperoxidase and eosinophil peroxidase.
Myeloperoxidase (MPO) and eosinophil peroxidase (EPO) are enzymes present in neutrophil and eosinophil leukocytes, respectively. Here, we present the development of a sensitive and specific assay for determination of the halogenating enzymatic activity of MPO and EPO based on the electrophilic attack of HOCl and HOBr on aromatic ring of dansylglycine (DG). We found that the intrinsic fluorescence of DG was promptly depleted by the action of these acids. In the presence of the enzymes, the fluorescence bleaching was dependent of chloride (Cl-) and bromide (Br-), which makes the assay able to distinguish the halogenating from the peroxidase activity. A linear correlation was obtained between the hydrogen peroxide (H2O2) concentration and the fluorescent decay. Similarly, the enzyme activity was measured by keeping constant H2O2. The method was applied for studding MPO/EPO specific inhibitors as 5-fluortryptamine (reversible inhibitor) and 4-hydroxybenzhydrazide (irreversible inhibitor). Differently of the taurine chloramine/3,3',5,5'-tetramethylbenzidine assay, which is among the most used technique, the dansylglycine assay was able to differentiate these inhibitors based on their kinetic behavior. In conclusion, this assay can differentiate the peroxidase and halogenating activity of MPO and EPO. Moreover, the method is adequate for real-time measurement of the production of HOCl and HOBr.


Beneficial effect of an inhibitor of leukocyte elastase (EPI-hNE-4) in presence of repeated lung injuries.
Persistence of alveolar neutrophil influx and activation may enhance the fibrotic process after acute lung injury. On the other hand, elastase has an antimicrobial activity and could participate in neutrophil migration, both events being critically important in host defense, explaining the controversial issue of therapeutic elastase inhibition in the setting of acute lung injury. We assessed the effect of a neutrophil elastase inhibitor, EPI-hNE-4, in single (bleomycin, 1.2 mg/rat intratracheally) and repeated (bleomycin, 1.2 mg/rat plus endotoxin and 1 mg/kg intratracheally 24 h later) lung injuries to assess the role of neutrophil in fibrosis. Subsequently, the effect of EPI-hNE-4 on bacterial clearance was evaluated during Pseudomonas aeruginosa-induced pneumonia. In the single injury model, despite a dramatic reduction of alveolar neutrophil influx with EPI-hNE-4 treatment, no significant inhibition of the decrease in respiratory system compliance, an index of lung fibrosis, was demonstrated at day 14. In the repeated injury model, EPI-hNE-4 treatment afforded a significant protective effect on compliance and alveolar inflammation at day 14. During bacterial pneumonia, EPI-hNE4 did not modify alveolar neutrophil recruitment or bacterial clearance from bronchoalveolar lavage fluid and lung homogenate. In conclusion, EPI-hNE-4, a specific inhibitor of leukocyte elastase, afforded a partial protective effect on the respiratory system compliance during repeated lung injuries, and had no detrimental effect during a gram-negative bacterial pneumonia.


A case of cerebrospinal fluid eosinophilia associated with shunt malfunction.
A 3-month-old female patient presented with a meningomyelocele at the lumber region associated with congenital hydrocephalus. She underwent ventriculoperitoneal (V-P) shunt surgery using the Sophy system. The shunt system was replaced due to a malformation. Following replacement, the patient presented with cerebrospinal fluid (CSF) eosinophilia at the age of 8 months. The eosinophilic granulocytosis of the CSF improved dramatically following systemic prednisolone administration. CSF eosinophilia without accompanying inflammation or pyrexia in the present case may have resulted from an allergic response to a foreign material such as the silicone tube pressure valve of the Sophy system or the sutures rather than bacterial or fungal infection. Based on our results, we believe that some patients may experience CSF eosinophilia following postoperative V-P shunt due to an allergic reaction to the shunt equipment. Prompt steroid treatment can produce spontaneous regression in such cases.


[Idiopathic recurrent facial palsy].
We report a 34 year old female patient with six episodes of idiopathic right facial palsy, all of wich had total recovery. Idiopathic recurrent facial palsy is a rare entity, with a few cases in the literature. Recurrent facial palsy is usually in the setting of a family history and have both autosomal dominant and recessive inheritance or in the setting of hypertension and diabetes. A review of the literature does not report a previous case with six episodes.


Proteomic identification of sorting nexin 6 as a negative regulator of BACE1-mediated APP processing.
The beta-site APP cleaving enzyme-1 (BACE1) mediates the first cleavage of the beta-amyloid precursor protein (APP) to yield the amyloid beta-peptide (Abeta), a key pathogenic agent in Alzheimer's disease (AD). Using a proteomic approach based on in-cell chemical cross-linking and tandem affinity purification (TAP), we herein identify sorting nexin 6 (SNX6) as a BACE1-associated protein. SNX6, a PX domain protein, is a putative component of retromer, a multiprotein cargo complex that mediates the retrograde trafficking of the cation-independent mannose-6-phosphate receptor (CI-MPR) and sortilin. RNA interference suppression of SNX6 increased BACE1-dependent secretion of soluble APP (sAPPbeta) and cell-associated fragments (C99), resulting in increased Abeta secretion. Furthermore, SNX6 reduction led to elevated steady-state BACE1 levels as well as increased retrograde transport of BACE1 in the endocytic pathway, suggesting that SNX6 modulates the retrograde trafficking and basal levels of BACE1, thereby regulating BACE1-mediated APP processing and Abeta biogenesis. Our study identifies a novel cellular pathway by which SNX6 negatively modulates BACE1-mediated cleavage of APP.


Sperm motility-activating complex formed by t-complex distorters.
Transmission ratio distortion is a dramatic example of non-Mendelian transmission. In mice, t-haplotype males produce dysfunctional +-sperm and normal t-sperm, leading to transmission in favor of t-sperm. Genetic studies have indicated that the t-complex responder locus, Tcr, rescues t-sperm but not +-sperm from defective products of t-complex distorter loci, Tcds. Light chain 1 (LC1) and LC3 from sea urchin sperm outer arm dynein have sequence similarities to Tctex2 and Tctex1, respectively, both of which are wild-type products of Tcds. We show here that LC1 and LC3 are able to make a 1:1 complex. Since Tcr is a member of the Smok (sperm motility kinase) family and LC1 is phosphorylated at the activation of sperm motility in a cAMP-dependent manner, this complex in a dynein motor molecule might be a direct target of Smok/Tcr kinase in a signal cascade that regulates sperm motility. Thus, we designate it as Smoac (sperm motility activating complex).


Silibinin and ellagic acid increase the expression of insulin receptor substrate 1 protein in ultraviolet irradiated rat skin.
Daily exposure to ultraviolet (UV) light induces inflammation and tumorigenesis in the skin. Silibinin and ellagic acid are natural products that exhibit anti-inflammatory and anti-tumorigenic properties. Insulin receptor substrate protein 1 (IRS1) is important for skin homeostasis and physiology, but its activity following UV radiation remains unclear. We investigated the effects of ellagic acid and silibinin on IRS1 expression in ultraviolet A (UVA) and ultraviolet B (UVB) irradiated rat skin. Forty-two female Wistar rats were divided randomly into six groups of seven animals. The dorsal skin of rats was exposed to UVA + UVB, then treated with ellagic acid and silibinin by gavage. IRS1 expression in skin tissues was determined by western blot analysis. IRS1 expression increased significantly following treatment with ellagic acid and silibinin in UVA + UVB irradiated skin compared to the UVA + UVB only group. After UVA + UVB treatment, ellagic acid effected greater induction of IRS1 expression than silibinin. Our findings suggest that the photoprotective roles of ellagic acid and silibinin may be due to induction of IRS1 expression in UVA + UVB treated rat skin.


Quantifying psychiatric comorbidity--lessions from chronic disease epidemiology.
Comorbidity research in psychiatric epidemiology mostly uses measures of association like odds or risk ratios to express how strongly disorders are linked. In contrast, chronic disease epidemiologists increasingly use measures of clustering, like multimorbidity (cluster) coefficients, to study comorbidity. This article compares measures of association and clustering. Narrative review, algebraical examples, a secondary analysis of an existing dataset and a pooled analysis of published data. Odds and risk ratios, but the former more than the latter, confound clustering with coincidental comorbidity. Multimorbidity coefficients provide a pure estimate of clustering which is the proportion of the association between disorders that is of etiological interest. Odds and risk ratios can express comorbidity between no more than two disorders, whilst clustering coefficients, although computationally laboursome, can capture multimorbidity of any number of disorders. Cluster coefficients depend less on the prevalence of illness in study groups than measures of association. Odds and risk ratios are well suited for comorbidity research which focuses on which sets of disorders or syndromes tend to occur in combination and the implications of this for, for instance, nosological classification, a traditional interest of psychiatric epidemiology. However, the cluster coefficient is to be preferred if the interest is more aetiological, addressing for example why certain individuals are prone to multiple health problems.


Cortex controls multisensory depression in superior colliculus.
Multisensory depression is a fundamental index of multisensory integration in superior colliculus (SC) neurons. It is initiated when one sensory stimulus (auditory) located outside its modality-specific receptive field degrades or eliminates the neuron's responses to another sensory stimulus (visual) presented within its modality-specific receptive field. The present experiments demonstrate that the capacity of SC neurons to engage in multisensory depression is strongly dependent on influences from two cortical areas (the anterior ectosylvian and rostral lateral suprasylvian sulci). When these cortices are deactivated, the ability of SC neurons to synthesize visual-auditory inputs in this way is compromised; multisensory responses are disinhibited, becoming more vigorous and in some cases indistinguishable from responses to the visual stimulus alone. Although obtaining a more robust multisensory SC response when cortex is nonfunctional than when it is functional may seem paradoxical, these data may help explain previous observations that the loss of these cortical influences permits visual orientation behavior in the presence of a normally disruptive auditory stimulus.


Differential expression patterns of two new primary cell wall-related cellulose synthase cDNAs, PtrCesA6 and PtrCesA7 from aspen trees.
Based on elegant molecular genetic analyses, distinct classes of cellulose synthase (CesA) genes have been associated with either primary or secondary cell wall development in Arabidopsis. Here, we report on cloning of two new CesA cDNAs, PtrCesA6 and PtrCesA7 involved in the primary cell wall development in aspen (Populus tremuloides) trees. Both these distinct cDNAs, isolated from a developing xylem cDNA library, share only 60-67% identities with each other as well as with five other previously known aspen CesA cDNAs. Interestingly, PtrCESA6 from aspen, a dicot species, shares maximum identity of 81-84% with three CESA isoforms from maize and rice, two monocot species. On the other hand, PtrCESA7 shares a maximum identity of 86% with AtCESA2, a primary wall-related CesA member from Arabidopsis, a dicot species. Gene expression analyses by reverse transcriptase-polymerase chain reactions (RT-PCRs) suggested that both these genes are expressed at a low level in all aspen tissues examined but PtrCesA7 is expressed at a higher level than PtrCesA6. While corroborating these results, in situ mRNA hybridization studies using three different aspen organs also suggested that PtrCesA6 and PtrCesA7 genes are expressed in all expanding cells depositing primary cell wall but PtrCesA7 is expressed at a higher level than PtrCesA6. These differential gene expression profiles suggest that each of these CesAs may be playing a specific role during primary cell wall development in aspen trees. Isolation of two primary wall related CesA genes from xylem tissues also suggest their importance during xylem development, which is traditionally considered to be enriched in secondary cell wall forming cells of economical significance.


Overcoming Difficulties in Molecular Biological Analysis through a Combination of Genetic Engineering, Genome Editing, and Genome Analysis in Hexaploid Chrysanthemum morifolium.
Chrysanthemum is one of the most commercially important ornamental plants globally, of which many new varieties are produced annually. Among these new varieties, many are the result of crossbreeding, while some are the result of mutation breeding. Recent advances in gene and genome sequencing technology have raised expectations about the use of biotechnology and genome breeding to efficiently breed new varieties. However, some features of chrysanthemum complicate molecular biological analysis. For example, chrysanthemum is a hexaploid hyperploid plant with a large genome, while its genome is heterogeneous because of the difficulty of obtaining pure lines due to self-incompatibility. Despite these difficulties, an increased number of reports on transcriptome analysis in chrysanthemum have been published as a result of recent technological advances in gene sequencing, which should deepen our understanding of the properties of these plants. In this review, we discuss recent studies using gene engineering, genome editing, and genome analysis, including transcriptome analysis, to analyze chrysanthemum, as well as the current status of and future prospects for chrysanthemum.


The Co-Occurrence of Conduct Problems and Depressive Symptoms From Childhood to Adulthood for Men: Stability Over Time and Prediction to Substance Use.
The dual pathway hypothesis of risk for substance use was tested by examining risk from symptoms of conduct problems and depressive symptoms in adolescence (from ages 10-11 to 17-18 years) to substance use-including tobacco, alcohol, cannabis, and other illicit drugs-in both early adulthood (approximately from ages 20 to 29 years) and middle adulthood (approximately from ages 29 to 38 years). Hypotheses were tested on a sample of boys who were at risk for conduct problems by virtue of the neighborhoods where they lived in childhood (the Oregon Youth Study; N = 206 at Wave 1). Dual-trajectory modeling (Latent Class Analysis) resulted in a 3-group solution of high, moderate, and low co-occurring symptoms. The latent class of boys with co-occurring symptoms in adolescence showed higher levels of substance use in adulthood; namely, higher levels of cannabis and illicit substance use during early adulthood compared to either of the moderate or low symptom classes, and higher use of cannabis in midadulthood than the low symptom class. Those with co-occurring symptoms also showed, overall, higher vulnerability to use of tobacco in these 2 periods, but not to higher use of alcohol. Regression analyses indicated that the higher substance use of the co-occur group of men was related to their adolescent conduct problems, but was not related to their adolescent depressive symptoms; however, these associations were nonsignificant when adolescent use of the respective substances were included in the models. Thus, the dual-trajectory hypothesis was not supported. However, the findings indicated that, as assessed in the present study, the psychopathology symptoms of boys with conduct problems in adolescence who show risk for later substance use may be complex, involving depressive symptoms.


[Intrahepatic cholestasis of pregnancy].
The review of intrahepatic cholestasis of pregnancy attempts to summarize the current knowledge of this disease by analysing available literary sources. Intrahepatic cholestasis of pregnancy is a disease that typically appears in the third trimester of pregnancy, sometimes already at the end of the second trimester of pregnancy. The main symptom of the disease is pruritus. In addition, the disease is characterized by increased levels of liver enzymes and bile acids. The symptoms of the disease disappear spontaneously after delivery. The disease is associated with high incidence of fetal distress, as well as with a high risk of premature labour. The most serious obstetric complication is antenatal sudden fetal death. Fetal complications are probably caused by elevated levels of bile acids. Therefore the aim of treatment should be to minimize negative effects of bile acids on the fetus, to prolong pregnancy and reduce maternal symptoms at the same time.


Routine pre-exposure rabies prophylaxis: a reassessment.
The rational use of rabies biologics is a difficult challenge. Pre-exposure rabies prophylaxis (PRE-RP) is one important rabies prevention strategy currently recommended by the US Public Health Service for groups considered at high risk of exposure to rabies. However, standard post-exposure rabies prophylaxis (POST-RP) appears equally effective in preventing rabies; PRE-RP still requires a limited vaccination series if a rabies exposure occurs and, at current costs, standard POST-RP may be a less expensive strategy than PRE-RP. For PRE-RP, as with other human rabies prevention strategies, the central problem is to obtain reasonable estimates of actual rabies exposure risks, which provide the rationale for well-designed programs.


Conditions for autonomous choice: a qualitative study of older adults' experience of decision-making in TAVR.
Patient autonomy is a leading principle in bioethics and a basis for shared decision making. This study explores conditions for an autonomous choice experienced by older adults who recently underwent trans-catheter aortic valve replacement (TAVR). Qualitative study entailing semi-structured interviews of a purposive sample of ten older (range 73-89, median 83.5 years) adults after TAVR (median 23 days). The study setting was a cardiac department at a university hospital performing TAVR since 2010. Analysis was by systematic text condensation. Even when choice seemed hard or absent, TAVR-patients deliberately took the chance offered them by processing risk assessment, ambivalence and fate. They regarded declining the treatment to be worse than accepting the risk related to the procedure. The experience of being thoroughly advised by their physician formed the basis of an autonomous trust. The trust they felt for the physicians' recommendations mitigated ambivalence about the procedure and risks. TAVR patients expressed feelings consistent with self-empowerment and claimed that it had to be their decision. Even so, choosing the intervention as an obligation to their family or passively accepting it was also reported. Older TAVR patients' experience of an autonomous decision may encompass frank tradeoff; deliberate physician dependency as well as a resilient self-view. Physicians should be especially aware of how older adults' subtle cognitive declines and inclinations to preserve their identities which can influence their medical decision making when obtaining informed consent. Cardiologists and other providers may also use these insights to develop new strategies that better respond to such inherent complexities.


Effects of insulin and glucagon on energy and carbohydrate metabolism of rat hepatocytes in primary culture.
We studied the effects of insulin and glucagon on energy and carbohydrate metabolism of rat hepatocytes in primary culture. The aim of this study is to elucidate the mechanism of the synergistic action of insulin and glucagon and to evaluate the combined effects of these hormones on liver injury. Insulin increased the level of adenosine triphosphate in hepatocytes in the presence of glucagon. Insulin increased the activities of glucokinase (EC 184.108.40.206), phosphofructokinase (EC 188.8.131.52), pyruvate kinase (EC 18.104.22.168) type L and glucose 6-phosphate dehydrogenase (EC 188.8.131.52). Glucagon had no antagonistic effect on these increases. Glucagon increased the activity of glucose 6-phosphate (EC 184.108.40.206) (G6Pase) in the presence or absence of insulin, while insulin had no effects on the levels of G6Pase and fructose 1,6-bisphosphatase (EC 126.96.36.199) in the presence or absence of glucagon. Metabolite analysis of cultured hepatocytes indicated that insulin and glucagon have antagonistic effects on the glycolytic activity of hepatocytes. These combined effects of insulin and glucagon may partially explain the preventive effects of these hormones on liver injury.


Differences of LAK-activity and IL-2 responsiveness between alpha/beta and gamma/delta T cells which developed after thymus transplantation.
A patient with severe combined immunodeficiency was transplanted with T cell depleted haploidentical bone marrow from his father and was later given a thymic graft from an unrelated donor. alpha/beta and gamma/delta T cells of bone marrow donor origin appeared only after the thymus transplantation procedure. Among the peripheral blood lymphocytes (PBL), gamma/delta T cells comprised 10-20% and most of them were delta TCS1+. The alpha/beta T cells were single positive cells, either CD4+ or CD8+. Expression of CD5, CD7 and CD8 alpha,beta molecules on alpha/beta T cells was reduced. Functional studies showed that gamma/delta T cells proliferated slightly in response to anti-CD3 stimulation, and proliferated well with exogenous IL-2 stimulation, while alpha/beta T cells did not proliferate following mitogenic stimulation even in the presence of IL-2. gamma/delta T cells but not alpha/beta T cells exhibited some LAK activity after culturing with IL-2. Since alpha/beta T cells expressed IL-2R alpha and beta chains after mitogenic stimulation and bound IL-2, the deficit(s) in these cells was considered to occur after IL-2 binding to the IL-2R. These results indicate thymic dependency of both types of T cells and that two types of T cells differed in the acquisition of IL-2 responsiveness during development.


Small-cell-type poorly differentiated squamous cell carcinoma of the lung. Cytologic, immunohistochemical and nuclear DNA content analysis.
The relationship between the nuclear DNA content, the immunohistochemical findings, the clinical characteristics (tumor volume doubling time and survival) and the cytomorphologic features of small cell poorly differentiated squamous cell carcinoma of the lung was studied in ten cases. There were no significant correlations between the immunohistochemical stainings for neuron-specific enolase and keratin and the clinical characteristics in these cases. The DNA histogram patterns were classified as type I or type II, depending on the degree of dispersion of values. There was no relationship between the immunohistochemical findings and the DNA histogram patterns. Only the DNA histogram patterns were related to some of the clinical characteristics: patients with type II histograms had significantly shorter tumor volume doubling times than did patients with type I histograms. Such information may aid in distinguishing the small cell type of poorly differentiated squamous carcinoma from classic small cell carcinoma of the lung, with which it may be confused.


Homologous and crossreacting immune response of the jird and cotton rat against microfilariae of Dipetalonema viteae and Litomosoides carinii (Nematoda: Filarioidea).
Consecutive injections of microfilariae of L. carinii and D. viteae were carried out in the homologous or heterologous sequence into the corresponding natural or into the experimental hosts. Microfilariae of D. viteae are imunogenic in Sigmodon specifically and crossreacting as well. However, in the natural host, Meriones, they are doubtlessly tolerogenic. The sojourn and level of microfilaraemia are prolonged resp. higher when living D. viteae-Mf are injected s.c. followed by an i.v. injection. However, freeze-killed microfilariae of D. viteae are immunogenic in Meriones. Obviously the two filarial worms living in small mammals realize the balance with their natural hosts in quite a different manner. Which one of the two possibilities, or if one of them at all, reflects the relationship in human pathogenic filariae remains open.


A Bioinspired Hierarchical Fast Transport Network Boosting Electrochemical Performance of 3D Printed Electrodes.
Current 3D printed electrodes suffer from insufficient multiscale transport speed, which limits the improvement of electrochemical performance of 3D printed electrodes. Herein, a bioinspired hierarchical fast transport network (HFTN) in a 3D printed reduced graphene oxide/carbon nanotube (3DP GC) electrode demonstrating superior electrochemical performance is constructed. Theoretical calculations reveal that the HFTN of the 3DP GC electrode increases the ion transport rate by more than 50 times and 36 times compared with those of the bulk GC electrode and traditional 3DP GC (T-3DP GC) electrode, respectively. Compared with carbon paper, carbon cloth, bulk GC electrode, and T-3DP GC electrode, the HFTN in 3DP GC electrode endows obvious advantages: i) efficient utilization of surface area for uniform catalysts dispersion during electrochemical deposition; ii) efficient utilization of catalysts enables the high mass activity of catalysts and low overpotential of electrode in electrocatalytic reaction. The cell of 3DP GC/Ni-NiO||3DP GC/NiS2 demonstrates a low voltage of only 1.42 V to reach 10 mA cm-2 and good stability up to 20 h for water splitting in alkaline conditions, which is superior to commercialized Pt/C||RuO2 . This work demonstrates great potential in developing high-performance 3D printed electrodes for electrochemical energy conversion and storage.


Endoscopic approach for diagnosing autoimmune pancreatitis.
It is of utmost importance to differentiate autoimmune pancreatitis (AIP) from pancreatic cancer (PC). Segmental AIP cases are sometimes difficult to differentiate from PC. On endoscopic retrograde cholangiopancreatography, long or skipped irregular narrowing of the main pancreatic duct (MPD), less upstream dilatation of the distal MPD, side branches derived from the narrowed portion of the MPD, absence of obstruction of the MPD, and stenosis of the intrahepatic bile duct suggest AIP rather than PC. Abundant infiltration of IgG4-positive plasma cells is frequently and rather specifically detected in the major duodenal papilla of AIP patients. IgG4-immunostaining of biopsy specimens obtained from the major duodenal papilla is useful for supporting a diagnosis of AIP with pancreatic head involvement. On endoscopic ultrasonography (EUS), hyperechoic spots in the hypoechoic mass and the duct-penetrating sign suggest AIP rather than PC. EUS and intraductal ultrasonography sometimes show wall thickening of the common bile duct even in the segment in which abnormalities are not clearly observed with cholangiography in AIP patients. EUS-guided fine needle aspiration, especially EUS-guided Tru-Cut biopsy, is useful to diagnose AIP, as well as to exclude PC.


Role of MaABI5-like in abscisic acid-induced cold tolerance of 'Fenjiao' banana fruit.
Abscisic acid (ABA) is a phytohormone essential for plants to respond to various environmental stresses, and abscisic acid-insensitive 5 (ABI5) is a basic leucine zipper transcription factor of the ABA signaling pathway. Exogenous ABA induces cold tolerance in bananas; however, the role of MaABI5-like in ABA-induced cold tolerance remains unexplored. The present study found that exogenous ABA alleviated chilling injury of 'Fenjiao' banana, induced the accumulation of endogenous ABA, unsaturated fatty acids, and flavonoid content, and reduced the saturated fatty acid content. Moreover, ABA treatment upregulated the transcription levels of MaABI5-like, fatty acid desaturation genes, and flavonoid synthesis-related genes during cold storage. More interestingly, MaABI5-like directly interacted with the promoter of genes related to fatty acid desaturation (MaFAD3-1, MaFAD3-4, MaFAD3-5, MaFAD6-2, MaFAD6-3) and flavonoid synthesis (MaPAL-like, MaPAL-like1, MaC4H-like3, Ma4CL-like1, Ma4CL-like10, MaCHS6-4-like, and MaFLS) and activated their expressions. Furthermore, the transient overexpression of MaABI5-like in 'Fenjiao' banana fruit and ectopic expression in tomato plants enhanced cold tolerance and upregulated fatty acid desaturation and flavonoid synthesis-related gene transcript levels. The reduced expression of MaABI5-like by virus-induced gene silencing in 'Fenjiao' banana increased chilling injury and downregulated the expression of fatty acid desaturation and flavonoid synthesis-related genes. Thus, the study indicates that MaABI5-like regulates ABA-induced cold tolerance by increasing unsaturated fatty acid and flavonoid content.


Shadow enhancers modulate distinct transcriptional parameters that differentially effect downstream patterning events.
Transcription in the early Drosophila blastoderm is coordinated by the collective action of hundreds of enhancers. Many genes are controlled by so-called 'shadow enhancers', which provide resilience to environment or genetic insult, allowing the embryo to robustly generate a precise transcriptional pattern. Emerging evidence suggests that many shadow enhancer pairs do not drive identical expression patterns, but the biological significance of this remains unclear. In this study, we characterize the shadow enhancer pair controlling the gene short gastrulation (sog). We removed either the intronic proximal enhancer or the upstream distal enhancer and monitored sog transcriptional kinetics. Notably, each enhancer differs in sog spatial expression, timing of activation and RNA Polymerase II loading rates. In addition, modeling of individual enhancer activities demonstrates that these enhancers integrate activation and repression signals differently. Whereas activation is due to the sum of the two enhancer activities, repression appears to depend on synergistic effects between enhancers. Finally, we examined the downstream signaling consequences resulting from the loss of either enhancer, and found changes in tissue patterning that can be explained by the differences in transcriptional kinetics measured.


Carbamyl phosphate synthetase III, an evolutionary intermediate in the transition between glutamine-dependent and ammonia-dependent carbamyl phosphate synthetases.
The amino acid sequence of carbamyl phosphate synthetase (CPS) III from liver of spiny dogfish shark Squalus acanthias was deduced from the nucleotide sequence of its cDNA. Alignment of the derived amino acid sequence of CPS III with sequences of rat and frog CPS I and hamster CPS II reveals a high degree of amino acid identity, indicating that CPS III shares the same common ancestral genes as CPSs I and II. All of the CPSs examined show a high conservation of sequences in the adenine nucleotide binding domains and in residues that have been implicated in catalysis. The active-site cysteine residue required for glutamine-dependent activity by CPS II is preserved in the sequence of CPS III. Nevertheless, analysis of the protein sequences indicates that CPS III is more closely related to CPS I than to CPS II. The structure of CPS III, which is composed of a single polypeptide, is consistent with the view that CPS III evolved by fusion of separate genes coding for the glutaminase and synthetase domains of the enzyme and, like other CPSs, the synthetase domain evolved by duplication and fusion of an ancestral kinase gene. These results, together with the recent finding that frog CPS I retains the active site cysteine residue in the glutaminase domain required for glutamine-dependent activity, indicate that other amino acid substitutions critical for glutamine-dependent activity preceded loss of this catalytic cysteine residue. The results described here together with earlier biochemical evidence support the view that acetylglutamate and glutamine-dependent CPS III found in invertebrates and fish species represents an intermediate in the evolution of ancestral glutamine-dependent CPS II toward the acetylglutamate and ammonia-dependent CPS I of ureotelic terrestrial vertebrates.


A Bowman-Birk trypsin inhibitor with antiproliferative activity from Hokkaido large black soybeans.
A trypsin inhibitor, with an N-terminal sequence highly homologous to those of 8-kDa Bowman-Birk trypsin inhibitors, was isolated from the seeds of Hokkaido large black soybeans. The trypsin inhibitor was unadsorbed on SP-Sepharose but adsorbed on DEAE-cellulose and Mono Q. It inhibited proliferation in breast cancer (MCF-7) cells and hepatoma (Hep G2) cells with an IC50 of 35 and 140 microM, respectively. The trypsin inhibitory activity of the inhibitor was completely preserved after exposure to temperatures up to 100 degrees C for 30 min and to the pH range 2-13 for the same duration. The trypsin inhibitor inhibited HIV-1 reverse transcriptase with an IC50 of 38 microM, but was devoid of antifungal activity toward Fusarium oxysporum and Mycosphaerella arachidicola.


BRD7 Stabilizes P53 via Dephosphorylation of MDM2 to Inhibit Tumor Growth in Breast Cancer Harboring Wild-type P53.
Bromodomain-containing protein 7 (BRD7) was found to be down-expressed in nasopharyngeal carcinoma as well as breast cancer and to function as a potential tumor suppressor. BRD7 interacts with p53 and is required for p53-dependent oncogene-induced senescence. However, the mechanism how BRD7 functions as tumor suppressor roles in breast cancer remains unclear. MTT, colony formation assay, cell cycle, cell apoptosis, and tumorigenicity assays were performed to evaluate the biological functions of BRD7 in breast cancer cells in vitro and in vivo. Real-time PCR, western blot, luciferase reporter gene assays, and co-immunoprecipitation were used to examine the gene expression, transcription activation and protein-protein interaction. We reported that BRD7 effectively suppressed cell proliferation and tumor growth in vitro and in vivo. In addition, BRD7 increased p53 protein stability through ubiquitin-dependent proteasome pathway and regulated the expression of p53 downstream target genes by activating its transcriptional activity in breast cancers harboring wild-type p53. Mechanistically, BRD7 decreased phosphorylation and activation of MDM2 via inactivating its upstream kinase AKT depending on the bromodomain of BRD7, therefore BRD7 significantly reduced the amounts of phosphorylated MDM2 binding with p53 eventually decreasing ubiquitination level of p53. Furthermore, silencing the expression of p53 at least partly reversed the inhibition effect of BRD7 on cell proliferation and tumor growth in vitro and in vivo. Our studies identify that BRD7 stabilizes p53 by inhibiting the phosphorylation of MDM2 via AKT pathway dependent on its bromodomain to function as a tumor suppressor in breast cancer harboring wild-type p53.


[Climacteric mask of endogenous depressions].
Eighty-six women of the menopausal age with neurotic and neurosis-like disturbances in the presence of the disrupted menstrual cycle were followed up at the Kazan psychoneurological outpatient centre. Out of the total number, 32 cases of masked depression in the framework of cyclothymia imitating climacteric disorders were ascertained. The author specifies the clinical characteristics of endogenic masked depression and the diagnostic criteria allowing its differentiation from climacteric neurosis.


Tecadenoson: a novel, selective A1 adenosine receptor agonist.
Tecadenoson is a novel selective A1 adenosine receptor agonist that is currently being evaluated for the conversion of paroxysmal supraventricular tachycardia (PSVT) to sinus rhythm. By selectively targeting the A1 receptor, tecadenoson may be associated with fewer adverse effects such as flushing, dyspnea, chest discomfort, and hypotension than adenosine, which is a nonselective agonist of all 4 adenosine receptors. Based on the results of phase I and phase II clinical trials, tecadenoson appears to be an effective agent for producing rapid and sustained conversion of PSVT to sinus rhythm. Additionally, the adverse effects that are typically attributed to adenosine's nonselective stimulation of the A2A, A2B, and A3 receptors appear to occur less frequently with the use of tecadenoson. Tecadenoson also appears to be associated with a lower incidence of atrial fibrillation following conversion of PSVT compared with the rates that have been associated with adenosine in the literature. A randomized, prospective trial will need to be conducted in the future to appropriately compare the safety and efficacy of tecadenoson and adenosine.


Predictive significance of magnetic resonance imaging at 4 months of adjusted age in infants after a perinatal neurologic insult.
The aim of this prospective study was to evaluate the predictive significance of magnetic resonance imaging (MRI) performed at 4 months of corrected age in 60 neonates after a perinatal neurologic insult. Follow-up ranged from 2 to 5 years of chronological age. MRI examination was normal in 10; isolated external hydrocephalus was found in 15 infants. Twenty-three of these infants developed normally. Focal or multifocal lesions were shown in 6 infants, of whom 2 developed normally. Diffuse brain involvement was present in 29 cases as atrophy (n = 18), leukomalacia (n = 5), basal ganglia lesions (n = 3), and delayed myelination (n = 3). All but 4 infants showed neurologic impairment. MRI performed at 4 months of adjusted age is of prognostic significance in neonates who suffer a moderate or mild neurologic insult.


Comparison of Continuous Femoral Nerve Block with and Without Combined Sciatic Nerve Block after Total Hip Arthroplasty: A Prospective Randomized Study.
In association with the growing interests in pain management, several modalities to control postoperative pain have been proposed and examined for the efficacy in the recent studies. Various modes of peripheral nerve block have been proposed and the effectiveness and safety have been examined for each of those techniques. We have described our clinical experiences, showing that continuous femoral nerve block could provide a satisfactory analgesic effect after total hip arthroplasty (THA) procedure. In this study, we compared the effectiveness and safety of continuous femoral nerve block with and without sciatic nerve blockade on pain control after THA. Forty patients scheduled for THA were included in the study and randomly divided into 2 groups. Postoperative analgesic measure was continuous femoral nerve block alone, while the identical regimen of continuous femoral nerve block was combined with sciatic nerve block. The amount of postoperative pain was evaluated in the immediate postoperative period, 6 hours, and 12 hours after surgery. Moreover, postoperative complications as well as requirement of supplemental analgesics during the initial 12 hours after surgery were reviewed in the patient record. The obtained study results showed that the supplemental sciatic nerve blockade provided no significant effect on arrival at the postoperative recovery room, while the NRS pain score was significantly reduced by the combined application of sciatic nerve blockade at 6 and 12 hours after surgery. In the investigation of postoperative analgesiarelated complications, no major complication was encountered without significant difference in complication rate between the groups.


Pharmacodynamics of mycophenolate mofetil after nonmyeloablative conditioning and unrelated donor hematopoietic cell transplantation.
The immunosuppressive drug mycophenolate mofetil (MMF) is used after nonmyeloablative hematopoietic cell transplantation (HCT); however, limited pharmacodynamic data are available. We evaluated plasma concentrations of mycophenolic acid (MPA), the active metabolite of MMF, and outcomes in 85 patients with hematologic malignancies conditioned with fludarabine and 2 Gy total body irradiation followed by HLA-matched unrelated-donor HCT and postgrafting cyclosporine and MMF. The first 38 patients received MMF 15 mg/kg twice daily; the next 47 patients received MMF 3 times daily. MPA pharmacokinetics were determined on days 7 and 21. Comparing the twice-daily and 3-times-daily MMF groups, the mean total MPA concentration steady state (Css) was 1.9 and 3.1 microg/mL; the unbound Css was 18 and 36 ng/mL, respectively (P < .001). Sixteen patients with a total MPA Css less than 3 microg/mL had low (< 50%) donor T-cell chimerism (P = .03), and 6 patients with MPA Css less than 2.5 microg/mL had graft rejection. An elevated unbound Css was associated with cytomegalovirus reactivation (P = .03). There were no significant associations between MPA pharmacokinetics and acute graft-versus-host disease (GVHD) or relapse. We conclude that increased MPA Css's predicted higher degrees of donor T-cell chimerism after unrelated donor nonmyeloablative HCT and suggest that targeting MPA Css's greater than 2.5 microg/mL could prevent graft rejection.


Economic and social impact of out-of-pocket expenditure on households of patients attending public hospitals.
We aimed to generate evidence on the social and economic impact of out-of-pocket expenses incurred by households on illness. We did a hospital-based cross-sectional study including a convenience sample of 374 inpatients and outpatients. The median illness expenditure was the same (₹62 500) for inpatients and outpatients. Of all respondents, 51.3% among the rural and 65.5% among the urban patients were employed before illness, but after illness only 24.4% among the rural and 23.4% among the urban patients remained in employment. The proportion of rural households of different socioeconomic categories that experienced decrease in expenditure on food, education and health, and those who had to sell land or cattle, and the education of whose children suffered was statistically significant. The proportion of indebted families in different socioeconomic classes was also statistically significant among both rural and urban patients. The lowest socioeconomic strata depended mostly upon the financial support of their friends to tide over the financial crisis of an illness. Our study shows that out-of-pocket expenses on healthcare are a burden not only for the poor but also the middle classes.