Pelargonidin ameliorates CCl4-induced liver fibrosis by suppressing the ROS-NLRP3-IL-1β axis via activating the Nrf2 pathway.
Liver fibrosis is a histological change that often occurs due to hepatic stellate cell (HSC) activation and excessive formation of an extracellular matrix in the liver. Pelargonidin (PEL) is a natural anthocyanidin existing in blueberries, berries, strawberries, and red radishes and has been demonstrated to possess health beneficial effects. Herein, we investigated the effect of PEL on liver fibrosis induced by CCl4 and hepatic stellate cells induced by transforming growth factor-β (TGF-β). We found that PEL administration prevented liver injury and liver fibrosis induced by CCl4 in a dose-dependent manner. Further data revealed that PEL increased liver nuclear factor E2-related factor 2 (Nrf2) and reduced liver oxidative stress and the expression levels of NLRP3, caspase-1 and IL-1β. In TGF-β-challenged HSCs (LX-2 cells), PEL effectively inhibited the LX-2 cell activation. In addition, the anti-fibrosis effects of PEL in LX-2 cells were abolished by Nrf2 knockdown. In summary, our study demonstrated that PEL ameliorated CCl4-induced liver fibrosis and HSC activation induced by TGF-β. The possible molecular mechanisms of PEL in liver fibrosis may be attributed to its suppression of ROS-NLRP3-IL-1β signaling by Nrf2 activation.


Purification and properties of the Rous sarcoma virus internal enhancer binding factor.
The internal enhancer binding factor (IBF) that specifically binds sequences within the gag gene internal enhancer of Rous sarcoma virus Schmidt-Ruppin A was purified to near homogeneity from BHK cells. The polypeptides that constituted IBF DNA-binding activity were identified by sodium dodecyl sulfate-polyacrylamide gel analysis. As isolated from BHK cells, IBF consisted of two different but related polypeptides. One (IBF alpha) had a molecular weight of 40,000; the other (IBF beta) had a molecular weight of 20,000 and appeared to be a proteolytic product of IBF alpha. The site within the gag gene to which IBF bounds in vitro (internal enhancer site 2; nucleotides 856 to 878 of the Rous sarcoma virus genome) were demonstrated to function as a cis-acting transcriptional stimulatory element both in vivo and in vitro. By using HeLa cell nuclear transcription extracts, purified IBF was found to function as a trans-acting transcription factor that stimulated transcription in vitro. Purified IBF was also demonstrated to be very similar to EBP20 (K. Carlberg, T. A. Ryden, and K. Beemon, J. Virol. 62:1617-1624, 1988), and it may well belong to the same family of DNA-binding proteins.


Diet, the endothelium and atherosclerosis.
Dietary factors have been implicated in the difference in the risk of cardiovascular disease observed between populations. However, clinical trials and mechanistic studies of dietary modification are difficult to undertake, and the data set on dietary intervention for cardiovascular disease is now much smaller than that for pharmacological interventions. Moreover, the precise mechanism by which dietary modification might alter the risk of cardiovascular disease is uncertain. In this issue of Clinical Science, Ambring and co-workers investigate the effect of a Mediterranean-inspired diet on endothelial function and lipid fractions.


First Report of Pseudomonas syringae pv. actinidiae, the Causal Agent of Bacterial Canker of Kiwifruit in Slovenia.
In May 2013, 20 plants in a production orchard of kiwifruit (Actinidia deliciosa) cv. Hayward in the seaside area of Primorska showed small, angular, coalescing necrotic leaf spots and cankers on green shoots. In the following 2 weeks, disease progressed to wilting and shoot dieback with exudates. Symptoms were consistent with Pseudomonas syringae pv. actinidiae. Circular, flat, granulated colonies with entire margins were isolated from leaf spots on King's medium B (KB) and on sucrose nutrient agar with boric acid, cephalexine, and cycloheximide. Strains were purified on KB and showed weak fluorescence upon a prolonged incubation (>10 days) and belonged to P. syringae LOPAT group Ia (+---+). DNA was extracted from strains and plant extracts with Chelex 100 resin and Bio-Nobile QuickPick Plant Kit (Turku, Finland), respectively. PCR products of expected sizes were generated by PCR assays (2,4) from all strains and plant extract, supporting the strains as being P. syringae pv. actinidiae. Two strains (NIB Z 1870 and 1871) were further identified by cytochrome C oxidase (negative), glucose metabolism (oxidative), aesculine (negative), and nitrate (negative). Their partial rpoD gene sequences (GenBank Accession Nos. KJ724117 and KJ724118) (3) were identical to the sequence of the P. syringae pv. actinidiae pathotype strain NCPPB 3739 (FN433222, 100% coverage) and to the sequence of P. syringae pv. theae at 96% coverage (FN433271). BOX-PCR fingerprinting and multilocus sequence analysis (MLSA) based on four housekeeping genes gapA (KJ733923 and KJ733924), gltA (KJ733925 and KJ733926), gyrB (KJ733927 and KJ733928), and rpoD identified both strains as biovar 3, a highly virulent biovar of P. syringae pv. actinidiae (5). The pathogenicity of the two strains was confirmed on four plants of A. deliciosa 'Hayward' for each strain. Six-month-old plants were sprayed on the abaxial sides of leaves with 30 ml cell suspension prepared from a 72-h-old culture of the appropriate strain (~8 × 106 CFU/ml in 0.01 M MgSO4), covered with plastic bags for 24 h, and incubated under high relative humidity (80%) with 14 h daylight and 24/21°C day/night temperature. Three positive and three negative control plants were inoculated with the Italian P. syringae pv. actinidiae virulent strain K9 (kindly provided by Dr. Gian Luca Bianchi of the Plant Health Service of Friuli Venezia Giulia region) and 0.01 M MgSO4, respectively. After 7 days, water-soaked brown spots with pale green halos were observed on all plants inoculated with bacteria. Re-isolated bacteria were identical to the original strains in their morphology, PCR products, and rpoD sequences. Negative control plants did not develop symptoms, and no growth was observed on media. This is the first laboratory confirmation of bacterial canker of kiwifruit in Slovenia. Visual inspections carried out by the plant health authorities in 2013 and laboratory analysis confirmed additional infection with P. syringae pv. actinidiae in a single, nearby orchard. The pest status of P. syringae pv. actinidiae in Slovenia is officially declared as present, subject to official control (1). References: (1) EPPO Reporting Service. Online publication: http://archives.eppo.int/EPPOReporting/2014/Rse-1402.pdf . No. 02 2014/026, 2014. (2) A. Gallelli et al. J. Plant Pathol 93:425, 2011. (3) N. Parkinson et al. Plant Pathol. 60:338, 2011. (4) J. Rees-George et al. Plant Pathol. 59:453, 2010. (5) J. L. Vanneste et al. Plant Dis. 97:708, 2013.


Retinal pigment epithelial cells are heterogeneous in their expression of MHC-II after stimulation with interferon-gamma.
The pattern of interferon-gamma-induced major histocompatibility complex Class II antigen expression was evaluated on the retinal pigment epithelium. Experiments were performed in vitro using explant cultures of aged and fetal human eyes and in vivo in albino rabbits. The human explants were stimulated with 50 U ml-1 interferon-gamma for 3 days prior to immunostaining for Class II. The rabbit eyes were subretinally injected in vivo with 50 microl of interferon-gamma (500 U ml-1) and analyzed immunohistochemically 3 days later. A heterogeneous pattern of Class II expression was present in the interferon-gamma-stimulated retinal pigment epithelial cells, in both the in vivo and the in vitro experiments. In aged human eyes the percent of Class-II positive cells was higher in the periphery than in the posterior pole (macular region) after interferon-gamma stimulation (P<0.01). No such difference was found in the fetal eyes. These data demonstrate that retinal pigment epithelial cells are heterogeneous in their response to interferon-gamma. The results are supportive of previous studies demonstrating the structural and proliferative heterogeneity of the retinal pigment epithelium. Together, these studies provide support for the possibility of functional retinal pigment epithelial heterogeneity.


Dentists' incomes, fees, practice costs, and the Economic Stabilization Act: 1952 to 1976. Bureau of Economic and Behavioral Research.
Throughout the 20-year period from 1952 to 1972, dentists' net incomes, adjusted for inflation, increased and provided greater monetary rewards to the practice of dentistry. Evidence discloses a close association between the purchasing power of dentists' net incomes and the general level of economic activity (measured by the GNP inconstant dollars). Dentists are not immune to temporary recessions. The period from 1972 to 1974, with the imposed fee guidelines of Nixon's Economic Stabilization Act, was unusual. With effective fee guidelines during the latter two phases coupled with freely fluctuating input prices, dentists were caught in an income squeeze. During 1972 to 1974, the purchasing power of dentists' net incomes fell at a rate of 10.1% per year as the price index of practice costs outpaced dentists' fees. After the Economic Stabilization Act expired in 1974, trends returned to normal. Dentists' net incomes increased at rates consistent with changes in the GNP and dentists' fees increased slightly faster than the "Price Index of Cost of Conducting a Dental Practice."


Lawyer control of internal scientific research to protect against products liability lawsuits. The Brown and Williamson documents.
To understand how attorneys for the tobacco industry in general, and Brown and Williamson Tobacco Corporation (B&W) in particular, have responded to the threat of products liability litigation arising from smoking-induced diseases. Documents from B&W, the British American Tobacco Company (BAT), and other tobacco interests provided by an anonymous source, obtained from Congress, or received from the private papers of a former BAT officer. All available materials, including confidential reports regarding research and internal memoranda exchanged between tobacco industry lawyers. The documents demonstrate that the tobacco industry in general, and B&W in particular, were very concerned about the threat of products liability lawsuits, and they illustrate some of the steps taken by lawyers at one company to avoid the discovery of documents that might be useful to a plaintiff in such a lawsuit. These steps included efforts to control the language of scientific discourse on issues related to smoking and health, to bring all potentially damaging internal scientific documents under attorney work product and attorney-client privilege to avoid discovery, to remove "deadwood" documents, and to insulate B&W from knowledge of potentially damaging scientific information from other BAT companies.


Telomere-to-centromere ratio of bovine clones, embryos, gametes, fetal cells, and adult cells.
In 1997, Dolly, the first animal cloned from an adult cell, was born. It was announced in 1999 that Dolly might be aging faster than normal because her telomeres were shorter than age-matched control sheep. Telomeres, a repeated DNA sequence located at the ends of linear chromosomes, allow for base pair loss during DNA replication. Telomere shortening acts as a "mitotic clock," leading to replicative senescence. By using whole cell lysate and slot-blot analysis, we determined the telomere-to-centromere ratio (T/C) for bovine gametes, embryos, fetal tissues (brain, heart, lung, kidney, uterus, ovary, and skin), adult donor cells, and cloned embryos. Our data indicates a consistency in T/C among the various fetal tissues. The T/C of sperm is significantly lower than in oocytes. The T/C decreases from the oocyte to the 2-8-cell stage embryo, increases dramatically at the morula stage, and decreases at the blastocyst stage. Our data shows no significant difference in T/C between cloned embryos and in vitro fertilized (IVF) embryos, but there is a significant difference between cloned embryos and adult donor cells. In conclusion, the enucleated bovine oocyte has the ability to reestablish the telomere length of adult somatic cell donor nuclei.


Recurrent intussusception. Risks and features.
To determine the risk factors associated with recurrent intussusception (RI) and to characterize the timing, features, and complications of RI. Retrospective chart review. Children's Hospital and Medical Center, Seattle, Wash. All patients with a diagnosis of intussusception who underwent barium enema as treatment for reduction between October 1, 1979 and December 31, 1990. Children with RI (N = 23, seven with two or more recurrent episodes) were classified as the case group; children with a single intussusception (N = 234), controls. There were no statistically significant differences in age, sex, race, symptoms, duration of symptoms, or results of the physical examination between the case group and controls. Reduction of the initial intussusception by a barium enema occurred in 96% of patients in the case group vs 62% of the controls (odds ratio, 13.50; 95% confidence intervals, 2.10 to 563.4; P = .003). Only one of 33 episodes of RI followed an operative reduction. In comparing the first recurrent episode with the initial episode, there was a significant reduction in the proportion of patients presenting with lethargy (0% vs 30%; Fisher Exact Test, P = .009) or blood in the stool (5% vs 52%; P < .002) and a drop in the median duration of symptoms from 20 to 6 hours. Recurrent intussusception cannot be predicted by presenting features or symptoms; operative reduction due to a failed reduction by a barium enema reduces the risk of RI; and patients with RI have fewer symptoms with a shorter duration.


[Subjective experience and quality of life in schizophrenia].
The term quality of life has been gradually introduced in the field of Psychiatry. This term tries to integrate not only the symptoms in Psychiatric patients, but also other aspects, like individual perceptions, personal expectations, degree of independence, etc., in order to achieve a global understanding of the patient. The are few studies on quality of life in schizophrenia, especially there is a lack of self assessment scales that the patients can fill out about themselves and their psychological environment. The Seville Quality of Life Questionnaire tries to evaluate the subjective experience of patients with schizophrenia about their illness. The Seville Quality of Life Questionnaire also checks the presence or absence of groups of patients with schizophrenia with a better or worst quality of life. We have assessed 222 subjects that meet ICD-10 criteria diagnostic for schizophrenia, with the Seville Quality of Life Questionnaire, the Positive and Negative Symptom Scale specific psychopathology scale and the AMDP system general psychopathology scale. Our results show two clearly differentiate groups: one with better (Group 2: 108 patients) and one with worse (Group 1: 81 patients) quality of life. However, 33 patients could not be included in any of the two groups. In the worse quality of life group the most prominent factors were: lack of cognitive grasp, perplexity, fear of loss of control, and contained hostility. These four factors have a great influence on the worse quality of life. When we compare the Seville Quality of Life Questionnaire data with several the Positive and Negative Symptom Scale scores, we saw that the group of worse quality of life presents higher scores on both Negative and Positive Symptoms of schizophrenia, and also in the Positive and Negative Symptom Scale general psychopathology scale. In the AMDP system general psychopathology scale we saw three groups of significant symptoms in the Seville Quality of Life Questionnaire: 1. Difficult relationships with the external world, 2. Difficulty to understand what it happening (loss of associations); and 3. Affective alterations (ambivalence).Finally, these data suggests that it is probable that the Seville Quality of Life Questionnaire could be able to build a typology of patients with schizophrenia more centred in their pathology that in their psychophatology.


A sandwich-ELISA for the diagnosis of Peste des petits ruminants (PPR) infection in small ruminants using anti-nucleocapsid protein monoclonal antibody.
A sandwich ELISA test using PPR specific monoclonal antibody (clone 4G6) to an epitope of nucleocapsid protein has been developed. The test uses polyclonal sera to capture the antigen from clinical samples (swabs and tissues). Captured antigens from clinical samples are detected using PPR specific monoclonal antibody. The test is specific to PPR as it failed to detect rinderpest vaccine virus (RBOK strain). Varieties of clinical samples originating from laboratory experiments (n = 231) and from field (n = 259) were employed to test the efficacy of sandwich-ELISA test. The test compared very well with an internationally accepted commercial Immune-capture ELISA kit, which uses biotinylated monoclonal antibody against the nucleocapsid protein. On a parallel testing using 490 clinical samples, 4G6 MAb based sandwich ELISA had an overall relative diagnostic specificity of 92.8% and diagnostic sensitivity of 88.9% compared to the commercial kit. The newly developed test is free from prozone phenomenon. PPR outbreaks from various parts of India have been confirmed using the test. Findings suggested that the newly developed ELISA is suitable for PPR diagnosis under field conditions.


Production of shotgun libraries using random amplification.
In the following report, thermal cycling coupled with random 10-mers as primers was used to construct randomly amplified shotgun libraries (RASLs). This approach allowed shotgun libraries to be constructed from nanogram quantities of input DNA. RASLs contained inserts from throughout a target genome in an unbiased fashion and did not appear to contain chimeric sequences. This protocol should be useful for shotgun sequencing the genomes of unculturable organisms and rapidly producing shotgun libraries from cosmids, fosmids, yeast artificial chromosomes (YACs), and bacterial artificial chromosomes (BACs).


Alternative scoring models of STOP-bang questionnaire improve specificity to detect undiagnosed obstructive sleep apnea.
Obstructive sleep apnea (OSA) is common among surgical patients. The STOP-Bang questionnaire is a validated screening tool with a high sensitivity. However, its moderate specificity may yield fairly high false positive rate. We hypothesized that the specific combinations of predicting factors in the STOP-Bang questionnaire would improve its specificity. After research ethics approval, consented patients were asked to complete the STOP-Bang questionnaire and then underwent sleep studies. The predictive performance of the STOP-Bang alternative scoring models was evaluated. Five hundred sixteen patients with complete data on the STOP-Bang questionnaire and polysomnography were reported. When the STOP-Bang score was ≥ 3 (any 3 positive items), the sensitivity and specificity for identifying moderate-severe OSA was 87% and 31%, respectively. The specificity for any 2 positive items from the 4 STOP questions plus BMI > 35 kg/m(2), male gender, or neck circumference > 40 cm for identifying moderate-severe OSA was 85%, 77%, and 79%, respectively. Compared with STOP-Bang score ≥ 3, the predicted probability for severe OSA of the specific combinations of STOP score ≥ 2 + male and STOP score ≥ 2 + BMI increased by 36% and 42%, respectively. For severe OSA, the specific combination of STOP score ≥ 2 + BMI + male demonstrated a specificity of 97% and 86% increase in predicted probability versus any 4 positive items of STOP-Bang questionnaire. The specific constellations of predictive factors improved the specificity of STOP-Bang questionnaire. For patients with STOP score ≥ 2, male gender, and BMI > 35 kg/m(2) were more predictive than age ≥ 50 and neck circumference > 40 cm.


Ferrous-activated nicotinamide adenine dinucleotide-linked dehydrogenase from a mutant of Escherichia coli capable of growth on 1, 2-propanediol.
A nicotinamide adenine dinucleotide-linked dehydrogenase has been partially purified from a mutant of Escherichia coli K-12 able to grow on l-1,2-propanediol as carbon and energy source. This enzyme catalyzes the dehydrogenation at carbon 1 of l-1,2-propanediol, glycerol, 1,3-propanediol, ethylene glycol, and ethyl alcohol. The purified protein requires added ferrous or managanous ions. The V(max) and the apparent K(m) for a given substrate vary with the particular metal used.


Psychiatric intervention in primary care for mothers whose schoolchildren have psychiatric disorder.
Psychiatric disorder in schoolchildren has been linked to increased general practice attendance rates. This increase may, in part, be a result of maternal stress focused on the disturbed child, and of a decrease in confidence in parenting. A study was undertaken to pilot the feasibility of a single session, psychiatric intervention in primary care for mothers of disturbed children and to examine uptake rates and reported immediate and long-term effects. Single psychiatric sessions by a child psychiatrist in general practice were offered to mothers of 26 schoolchildren. The schoolchildren (age range 7-12 years, mean nine years) were frequent attenders in general practice with physical symptoms, and were identified from research interviews carried out with a parent (usually their mother) as psychiatrically disordered. The main outcome measures were the mothers' ratings of helpfulness of the intervention; degree of behavioural, emotional or health problems in their children and confidence in managing them; the Rutter A parental behaviour questionnaire; and children's yearly general practice attendance rates. Sixteen mothers (62%) who were offered appointments attended for the intervention. Nine of the 14 who responded at three-month follow up (64%) reported that the intervention had been markedly or extremely helpful. The main areas of perceived improvement at both three months and at 18-24 months were in the child's behaviour, emotional and health problems, and in the mother's confidence in dealing with these. Mothers also found the specific advice discussed and the ability to talk to somebody about the problems helpful. Mothers were less likely to find the intervention extremely or markedly useful where the child had had previous psychiatric intervention. The mean yearly attendance rate for the whole group of 23 children (data missing for three) decreased from 6.5 consultations before the intervention to 2.8 afterwards; there was a non-significant trend for the drop in attendance to be more marked in the group whose mothers attended the intervention and who felt helped by it, than among the group of children whose mothers only reported finding the intervention slightly useful. Standardized child psychiatric interventions which may be used in the primary care setting appear acceptable and may be helpful to mothers in addressing psychiatric disorders associated with somatic presentation in their schoolchildren.


Altered tumor formation and evolutionary selection of genetic variants in the human MDM4 oncogene.
A large body of evidence strongly suggests that the p53 tumor suppressor pathway is central in reducing cancer frequency in vertebrates. The protein product of the haploinsufficient mouse double minute 2 (MDM2) oncogene binds to and inhibits the p53 protein. Recent studies of human genetic variants in p53 and MDM2 have shown that single nucleotide polymorphisms (SNPs) can affect p53 signaling, confer cancer risk, and suggest that the pathway is under evolutionary selective pressure (1-4). In this report, we analyze the haplotype structure of MDM4, a structural homolog of MDM2, in several different human populations. Unusual patterns of linkage disequilibrium (LD) in the haplotype distribution of MDM4 indicate the presence of candidate SNPs that may also modify the efficacy of the p53 pathway. Association studies in 5 different patient populations reveal that these SNPs in MDM4 confer an increased risk for, or early onset of, human breast and ovarian cancers in Ashkenazi Jewish and European cohorts, respectively. This report not only implicates MDM4 as a key regulator of tumorigenesis in the human breast and ovary, but also exploits for the first time evolutionary driven linkage disequilibrium as a means to select SNPs of p53 pathway genes that might be clinically relevant.


Characterization of skin surface and dermal microbiota in dogs with mast cell tumor.
The skin microbiota interacts with the host immune response to maintain the homeostasis. Changes in the skin microbiota are linked to the onset and the progression of several diseases, including tumors. We characterized the skin surface and dermal microbiota of 11 dogs affected by spontaneous mast cell tumor (MCT), using skin contralateral sites as intra-animal healthy controls. The microbial profile differed between healthy and tumor skin surfaces and dermis, demonstrating that the change in microbiota composition is related to the presence of MCT. The number of observed taxa between MCT and healthy skin surfaces was detected, showing a decrease in number and heterogeneity of taxa over the skin surface of MCT, at both inter- and intra-individual level. Preliminary data on bacterial population of MCT dermis, obtained only on three dogs, demonstrated an intra-individual reduction of taxa number when compared to the skin surface. Taxonomy reveals an increase of Firmicutes phylum and Corynebacteriaceae family in MCT skin surface when compared to the healthy contralateral. In conclusion, we demonstrate that microbial population of skin surface and dermis is related to mast cell tumor. Our study provides the basis for future investigations aiming to better define the interaction between mast cell tumors, microbiota and host immune response.


The impacts of pesticides on the health of farmers in Fasa, Iran.
With the growing global population, it is becoming increasingly difficult to ignore the demands for food and energy. A major portion of the food and energy is produced via agriculture and livestock activities. The objectives of this research are to gather information regarding the demographic features of farmers, previous poisoning, and the extent of farmers' knowledge in the use of pesticides and associated hazards in different counties and villages in Fasa, a city located in the Fars province of Iran. This cross-sectional study was carried out from March to July 2012 in the Nobandegan and Sheshdeh counties, and villages including Miandeh, Fedshkuyeh, Senan, Rahimabad, and other places in the Fasa suburban countryside. To collect data, an appropriate questionnaire was designed and implemented. A total of 200 farmers participated in the study. We found that 55% of farmers were illiterate. Approximately 86% of used pesticides were organophosphorus compounds. Around 30% of the farmers used no protective equipment while working with pesticides, and only 22% of farmers had read and understood the instructions on the pesticide containers. Given the toxicity and hazards of pesticides and their adverse effects on farmers' health, effective measures should be adopted to decrease the amount of pesticides used. Conducting training programs for farmers may help to reduce pesticide exposure risks.


Characteristics associated with purchasing antidepressant or antianxiety medications through primary care in Israel.
This study analyzed the role of patient and physician characteristics associated with the purchase of antidepressant or antianxiety medications in Israel, a country that has a universal health care system. A national sample of 30,000 primary care patients over the age of 22 was randomly drawn from the registry of the largest health care fund in Israel. Data concerning medication purchase between January and December 2006 were extracted. Physician and patient characteristics were merged with Israel's unique identification number. Multilevel analysis was conducted to identify patient- and physician-level predictors of medication purchase. Overall, 19% (N = 4,762) of the sample purchased antidepressant or antianxiety medications. Individuals with greater general medical and psychiatric comorbidity were more likely to purchase antidepressant or antianxiety medications. Older adults, women, those of higher socioeconomic status, and immigrants (with the exception of Jews born in Asia or Africa) were also more likely to purchase medications. Arabs and Jews born in Asia and Africa were less likely to purchase medications even after all other variables were accounted for. Physician characteristics were minimally associated with the purchase of medications. The findings demonstrate that despite universal health care access, there were variations by population groups. Educational efforts should target patients as well as physicians.


Age-Related Changes in Binaural Interaction at Brainstem Level.
Age-related hearing loss hampers the ability to understand speech in adverse listening conditions. This is attributed to a complex interaction of changes in the peripheral and central auditory system. One aspect that may deteriorate across the lifespan is binaural interaction. The present study investigates binaural interaction at the level of the auditory brainstem. It is hypothesized that brainstem binaural interaction deteriorates with advancing age. Forty-two subjects of various age participated in the study. Auditory brainstem responses (ABRs) were recorded using clicks and 500 Hz tone-bursts. ABRs were elicited by monaural right, monaural left, and binaural stimulation. Binaural interaction was investigated in two ways. First, grand averages of the binaural interaction component were computed for each age group. Second, wave V characteristics of the binaural ABR were compared with those of the summed left and right ABRs. Binaural interaction in the click ABR was demonstrated by shorter latencies and smaller amplitudes in the binaural compared with the summed monaural responses. For 500 Hz tone-burst ABR, no latency differences were found. However, amplitudes were significantly smaller in the binaural than summed monaural condition. An age-effect was found for 500 Hz tone-burst, but not for click ABR. Brainstem binaural interaction seems to decline with age. Interestingly, these changes seem to be stimulus-dependent.


Ctenocystoidea: new class of primitive echinoderms.
The oldest known "carpoids" of the echinoderm subphylum Homalozoa have been discovered in lower Middle Cambrian rocks of northern Utah. They were free-living benthonic animals characterized by a unique ctenoid feeding apparatus and flattened flexible theca with near bilateral symmetry. A new class, Ctenocystoidea, is proposed to include the new genus and new species, Ctenocystis utahensis.


Two-Dimensional Frontier-Based Viewpoint Generation for Exploring and Mapping Underwater Environments.
To autonomously explore complex underwater environments, it is convenient to develop motion planning strategies that do not depend on prior information. In this publication, we present a robotic exploration algorithm for autonomous underwater vehicles (AUVs) that is able to guide the robot so that it explores an unknown 2-dimensional (2D) environment. The algorithm is built upon view planning (VP) and frontier-based (FB) strategies. Traditional robotic exploration algorithms seek full coverage of the scene with data from only one sensor. If data coverage is required for multiple sensors, multiple exploration missions are required. Our approach has been designed to sense the environment achieving full coverage with data from two sensors in a single exploration mission: occupancy data from the profiling sonar, from which the shape of the environment is perceived, and optical data from the camera, to capture the details of the environment. This saves time and mission costs. The algorithm has been designed to be computationally efficient, so that it can run online in the AUV's onboard computer. In our approach, the environment is represented using a labeled quadtree occupancy map which, at the same time, is used to generate the viewpoints that guide the exploration. We have tested the algorithm in different environments through numerous experiments, which include sea operations using the Sparus II AUV and its sensor suite.


Analysis of the glycine binding domain of the NMDA receptor channel zeta 1 subunit.
In an attempt to examine glycine binding domain of the zeta 1 subunit of the mouse N-methyl-D-aspartate (NMDA) receptor channel, we constructed N-terminal or C-terminal deletion mutants of the zeta 1 subunit cDNA and expressed them in Spodoptera frugiperda cells using a baculovirus system. Analysis of binding of a glycine binding site antagonist, 5,7-[3(-3)H]dichlorokynurenate ([3H]DCKA) to the deleted zeta 1 mutants provided the first direct experimental evidence that the regions of N-terminal 282 and C-terminal 102 amino acid residues do not significantly affect glycine binding, and that both the region of approximately 260 amino acid residues preceding the putative transmembrane segment M1 and the region between the segments M3 and M4 are required to form the glycine binding domain.


Production of transgenic mice carrying the human hepatitis B virus or the human papillomavirus DNA sequences in Taiwan: analysis of physical structure and hereditary mode.
In order to produce suitable animal model systems for the analysis of possible synergistic interaction between hepatitis B virus (HBV) or the human papillomavirus (HPV) with other viral or chemical co-factors, we have produced transgenic mice carrying HBV, HPV16 or HPV18. Transfer of 3343 micro-injected one-cell (C57/BL6 X CBA) F1 embryos into pseudo-pregnant ICR recipients produced 181 pups. Seventeen of 151 of the pups analysed were found to be transgenics. Southern blot and slot blot analysis showed that the copy number of the transgenome in the transgenic mice ranged from 1-50 copies per cell. All the transgenomes analysed appeared to be in tandem repeats without detectable sequence rearrangements. The transgenome sequences were also stable on transmission to the F3 generation. On breeding, six of the ten transgenic mice analysed showed the Mendelian mode of inheritance with integration at single or multiple loci. Of the remaining four mice, three were mosaics and one was sterile. Analysis of the gene expression of the transgenic mice is currently in progress.


Knockdown of protein phosphatase 5 (PPP5C) suppresses the growth of leukemic cell line U937.
Serine/threonine protein phosphatase 5 (PPP5C) participates in multiple signaling pathways including cell cycle control and cell growth. PPP5C is involved in the progression of human breast cancer and hepatocellular carcinoma. However, its function in acute myelogenous leukemia (AML) remains unknown. In this study, we constructed a lentivirus system to knock down the expression level of PPP5C in leukemic cell line U937. Cell proliferation and cell cycles were assessed by MTT assay and flow cytometry respectively. Western blot was used to determine the level of caspase-3, PARP (poly ADP-ribose polymerase), CDK4 and CyclinD1. Knockdown of PPP5C suppressed the proliferation ability of U937 cells, and led to G0/G1 phase arrest, inducing cell apoptosis in U937 cells. The apoptosis of the U937 cells was associated with upregulating cleaved caspase-3 and PARP, and downregulating CDK4 and CyclinD1. In conclusions, PPP5C knockdown inhibits U937 cell proliferation and might be used as a potential therapeutic target for the treatment of leukemia.


Neuroactive substances specifically modulate rhythmic body contractions in the nerveless metazoon Tethya wilhelma (Demospongiae, Porifera).
Sponges (Porifera) are nerve- and muscleless metazoa, but display coordinated motor reactions. Therefore, they represent a valuable phylum to investigate coordination systems, which evolved in a hypothetical Urmetazoon prior to the central nervous system (CNS) of later metazoa. We have chosen the contractile and locomotive species Tethya wilhelma (Demospongiae, Hadromerida) as a model system for our research, using quantitative analysis based on digital time lapse imaging. In order to evaluate candidate coordination pathways, we extracorporeally tested a number of chemical messengers, agonists and antagonists known from chemical signalling pathways in animals with CNS. Sponge body contraction of T. wilhelma was induced by caffeine, glycine, serotonine, nitric oxide (NO) and extracellular cyclic adenosine monophosphate (cAMP). The induction by glycine and cAMP followed patterns varying from other substances. Induction by cAMP was delayed, while glycine lead to a bi-phasic contraction response. The frequency of the endogenous contraction rhythm of T. wilhelma was significantly decreased by adrenaline and NO, with the same tendency for cAMP and acetylcholine. In contrast, caffeine and glycine increased the contraction frequency. The endogenous rhythm appeared irregular during application of caffeine, adrenaline, NO and cAMP. Caffeine, glycine and NO attenuated the contraction amplitude. All effects on the endogenous rhythm were neutralised by the washout of the substances from the experimental reactor system. Our study demonstrates that a number of chemical messengers, agonists and antagonists induce contraction and/or modulate the endogenous contraction rhythm and amplitude of our nerveless model metazoon T. wilhelma. We conclude that a relatively complex system of chemical messengers regulates the contraction behaviour through auto- and paracrine signalling, which is presented in a hypothetical model. We assume that adrenergic, adenosynergic and glycinergic pathways, as well as pathways based on NO and extracellular cAMP are candidates for the regulation and timing of the endogenous contraction rhythm within pacemaker cells, while GABA, glutamate and serotonine are candidates for the direct coordination of the contractile cells.


Curettage and reconstruction by the sandwich technique for giant cell tumours around the knee.
To evaluate outcomes of 36 patients who underwent curettage, use of phenol, and reconstruction using the sandwich technique for giant cell tumour (GCT) of bone around the knee. 22 women and 14 men aged 19 to 46 (mean, 29.6) years underwent intralesional curettage, use of phenol, and reconstruction using the sandwich technique for GCT of the proximal tibia (n=23) or distal femur (n=13). Two of the cases were recurrences. Two, 18, and 16 tumours were classified as grade I, grade II, and grade III, respectively. Five of the grade III tumours were associated with an extra-articular pathological fracture. Patients underwent intralesional curettage, use of phenol, and reconstruction with allograft, gel foam, and cement (the sandwich technique). Pathological fractures were fixed with plates. Functional outcome was evaluated using the Musculoskeletal Tumor Society (MSTS) score. The mean follow-up period was 5 (2.5-11) years. The mean MSTS score was 27.7 out of 30 (standard deviation, 3; range, 16-30). One patient with a grade III tumour in the proximal tibia had a recurrence detected elsewhere after 3 years. Her MSTS score at 2 years was 26. No patient had malignant transformation. Intralesional curettage, use of phenol, and reconstruction with allograft, gel foam, and cement (the sandwich technique) for GCT of bone achieved good functional outcome and a low recurrence rate.


A U-Net Ensemble for breast lesion segmentation in DCE MRI.
Dynamic Contrast Enhanced Magnetic Resonance Imaging (DCE-MRI) has been recognized as an effective tool for Breast Cancer (BC) diagnosis. Automatic BC analysis from DCE-MRI depends on features extracted particularly from lesions, hence, lesions need to be accurately segmented as a prior step. Due to the time and experience required to manually segment lesions in 4D DCE-MRI, automating this task is expected to reduce the workload, reduce observer variability and improve diagnostic accuracy. In this paper we propose an automated method for breast lesion segmentation from DCE-MRI based on a U-Net framework. The contributions of this work are the proposal of a modified U-Net architecture and the analysis of the input DCE information. In that sense, we propose the use of an ensemble method combining three U-Net models, each using a different input combination, outperforming all individual methods and other existing approaches. For evaluation, we use a subset of 46 cases from the TCGA-BRCA dataset, a challenging and publicly available dataset not reported to date for this task. Due to the incomplete annotations provided, we complement them with the help of a radiologist in order to include secondary lesions that were not originally segmented. The proposed ensemble method obtains a mean Dice Similarity Coefficient (DSC) of 0.680 (0.802 for main lesions) which outperforms state-of-the art methods using the same dataset, demonstrating the effectiveness of our method considering the complexity of the dataset.


Effects of lysosomal collagenolytic enzymes, anti-inflammatory drugs and other substances on some properties of insoluble collagen.
1. An enzyme system present in a rat liver lysosome-rich fraction was found to liberate soluble hydroxyproline-containing products from insoluble collagen, with maximum activity at pH3.45. It was concluded that a form of cathepsin D was involved since synthetic substrates specific for trypsin were not hydrolysed. Collagenolysis was enhanced by thiol compounds and inhibited by Cu(2+) ions and the anti-inflammatory drugs phenylbutazone and ibufenac. 2. The possibility that behaviour of collagen and collagenolysis were modified by various substances, either by destruction of intramolecular and intermolecular bonds in tropocollagen or by electrostatic interactions, is discussed. Insoluble collagen was found to bind electrostatically to chondromucoprotein. This interaction was inhibited by some anti-inflammatory drugs. 3. Possible roles of the lysosomal collagenolytic enzyme system in experimental lathyrism in rats given penicillamine, and in erosion of cartilage in rheumatoid arthritis, are considered. 4. Collagenolysis in vivo, which may depend on complex interrelationships between collagen, chondromucoprotein and metal ions, is discussed in relation to possible effects, both harmful and beneficial, of anti-inflammatory drugs used in rheumatoid arthritis.


[Women with pelvic complaints and atypical varicose veins, varicose veins of the vulva and insufficiency of the pelvic veins; treatment with embolisation].
In three women, aged 34, 52 and 30 years, respectively, who suffered from chronic pelvic symptoms such as a heavy feeling and abdominal pain, atypical varicose veins were observed on the medial side of the upper thighs together with varicose veins of the vulva; these are indicators of insufficiency of the pelvic veins. The symptoms were reduced following embolisation of the insufficient pelvic veins. These symptoms are also known as the pelvic congestive syndrome. The exact correlation between insufficiency of the pelvic veins and the complaints mentioned above has not yet been determined, but it seems that in most patients the symptoms diminish or disappear following embolisation of the insufficient pelvic veins. When patients complain of chronic pelvic pain of unknown aetiology, one should look for atypical varices and vulval varices and consider pelvic-vein insufficiency as a possible cause.


Management of status asthmaticus in children.
Recent literature on paediatric status asthmaticus (PSA) confirms an increasing percentage of admissions to paediatric intensive care units. PSA is a medical emergency that can be fatal and needs careful and prompt intervention. The severity of PSA is mainly determined by clinical judgement of signs and symptoms. Peak flow measurements and serial lung function measurements are not reliable in PSA. Validated clinically useful instruments are lacking. The three main factors that are involved in the pathophysiology of PSA, bronchoconstriction, mucus plugging and airway inflammation need to be addressed to optimise treatment. Initial therapies include supplementation of oxygen, repetitive administration of rapid acting β2-agonists, inhaled anticholinergics in combination with systemic glucocorticosteroids and intravenous magnesium sulphate. Additional treatment modalities may include methylxanthines, DNase, ketamine, sodium bicarbonate, heliox, epinephrine, non-invasive respiratory support, mechanical ventilation and inhalational anaesthetics.


A mathematical model for mechanotransduction at the early steps of suture formation.
Growth and patterning of craniofacial sutures is subjected to the effects of mechanical stress. Mechanotransduction processes occurring at the margins of the sutures are not precisely understood. Here, we propose a simple theoretical model based on the orientation of collagen fibres within the suture in response to local stress. We demonstrate that fibre alignment generates an instability leading to the emergence of interdigitations. We confirm the appearance of this instability both analytically and numerically. To support our model, we use histology and synchrotron X-ray microtomography and reveal the fine structure of fibres within the sutural mesenchyme and their insertion into the bone. Furthermore, using a mouse model with impaired mechanotransduction, we show that the architecture of sutures is disturbed when forces are not interpreted properly. Finally, by studying the structure of sutures in the mouse, the rat, an actinopterygian (Polypterus bichir) and a placoderm (Compagopiscis croucheri), we show that bone deposition patterns during dermal bone growth are conserved within jawed vertebrates. In total, these results support the role of mechanical constraints in the growth and patterning of craniofacial sutures, a process that was probably effective at the emergence of gnathostomes, and provide new directions for the understanding of normal and pathological suture fusion.


Epileptogenic action of intraventricularly injected antimelatonin antibody.
In rats, intraventricularly injected antimelatonin antibody caused the appearance of transitory epileptiform abnormalities. Epileptic activity arose from and was limited to the cortical mantle of the hemisphere ipsilateral to the injection side. The occasional occurrence of lateralized seizures has also been observed. Control injection of saline, of the vehicle (rabbit serum), or of antibody saturated with melatonin induced flattening and desynchronization of the electroencephalogram but not epileptiform activity. Repeated antimelatonin injections caused reappearance of the same type of epileptic abnormalities that lasted slightly longer than the first time. Melatonin may play an inhibitory role in neuronal excitability.


EBV Reactivation in Transplant Recipients following SARS-CoV-2 Infection: A Retrospective Study.
Reactivation and primary infection with a high Epstein Barr Virus (EBV) DNA level in kidney transplant patients could cause severe complications, including the development of Post-Transplantation Lymphoproliferative Disease (PTLD). While in the general population the reactivation of EBV after SARS-CoV-2 infection has been reported, very few data are available in transplant recipients. Our retrospective study aimed to evaluate a possible EBV reactivation in kidney transplant patients following SARS-CoV-2 infection and a possible impairment of the immune system. In addition, the effects of changes in immunosuppressive therapy on EBV DNA reactivation and vaccination were also evaluated. A total of 166 kidney transplant patients followed at the Kidney-Pancreas Transplant Ambulatory Nephrology Unit at Padova University Hospital were retrospectively considered for an observation period of 6 months from January 2020 to April 2023. EBV DNA level was measured by Rt-PCR and evaluated 6 months before and after SARS-CoV-2 infection. Patients' serological states were established via quantification of anti-VCA and anti-EBNA (chemiluminescence). Patients' immune systems were characterized by CD4+/CD8+ lymphocyte ratio (flow cytometry). EBV DNA was reactivated in 50% of the 166 patients with COVID-19 who completed the study. Older patients with more severe forms of COVID-19 had higher EBV reactivation (p < 0.05). EBV reactivation significantly increased in patients with severe SARS-CoV-2 infection requiring hospitalization compared to patients managed at home (p < 0.001). CD4+/CD8+ lymphocyte ratio was reduced in patients with a younger age of transplant (p < 0.01) and on a higher dose of steroids (p < 0.01). The results of our study confirm the role of immunodepression, especially in recent transplant patients and those on high steroids, in EBV reactivation. These results combined with the few available in the literature might contribute to providing an optimal management of immunosuppressive treatment for these patients in order to obtain an immune state unfavorable to the activation of latent viruses, including EBV.


Ultrafast polarization bio-imaging based on coherent detection and time-stretch techniques.
Optical polarization imaging has played an important role in many biological and biomedical applications, as it provides a label-free and non-invasive detection scheme to reveal the polarization information of optical rotation, birefringence, and photoelasticity distribution inherent in biological samples. However, the imaging speeds of the previously demonstrated polarization imaging techniques were often limited by the slow frame rates of the arrayed imaging detectors, which usually run at frame rates of several hundred hertz. By combining the optical coherent detection of orthogonal polarizations and the optical time-stretch imaging technique, we achieved ultrafast polarization bio-imaging at an extremely fast record line scanning rate up to 100 MHz without averaging. We experimentally demonstrated the superior performance of our method by imaging three slices of different kinds of biological samples with the retrieved Jones matrix and polarization-sensitive information including birefringence and diattenuation. The proposed system in this paper may find potential applications for ultrafast polarization dynamics in living samples or some other advanced biomedical research.


Autonomic pathways in development of neural stimulation-induced gastric mucosal damage.
Gastric mucosal erosions can be induced by electrical stimulation of either vagus nerves (5 Hz, 5 V, 1 ms) or the paraventricular nucleus (PVN) of the hypothalamus (200 microA, 60 Hz, 100-microseconds pulse width). We have utilized various pharmacological and surgical interventions to determine the contributions of different components of the autonomic nervous system to the development of this neurally induced gastric damage in urethan-anesthetized Sprague-Dawley rats. In all experiments damage was assessed macroscopically and scored blindly on a 0 (normal) to 3 (severe) scale with samples sectioned for subsequent histological assessment of damage at the light microscopic level. Animals pretreated with either hexamethonium (30 mg/kg iv) or atropine (2 mg/kg iv) demonstrated reduced gastric damage scores after vagal stimulation compared with untreated control animals (P < 0.05). In contrast animals that underwent cervical cord transection exhibited gastric damage after both vagal and PVN stimulation that was not significantly different compared with animals with an intact cord undergoing similar stimulation (P > 0.05). Such cord transection itself did not cause any significant change to the gastric mucosa in the time period studied. These data emphasize the importance of the autonomic nervous system, in particular the parasympathetic component in the development of vagal stimulation-induced gastric damage. In addition, the present studies suggest that neither vagal nor PVN stimulation-induced gastric damage is dependent on neural projections to sympathetic preganglionic neurons of the intermediolateral cell column of the spinal cord.


Computational modeling and fluorescence microscopy characterization of a two-phase magnetophoretic microsystem for continuous-flow blood detoxification.
Magnetic beads can be functionalized to capture and separate target pathogens from blood for extracorporeal detoxification. The beads can be magnetically separated from a blood stream and collected into a coflowing buffer solution using a two-phase liquid-liquid continuous-flow microfluidic device in the presence of an external field. However, device design and process optimization, i.e. high bead recovery with minimum blood loss or dilution remain a substantial technological challenge. We introduce a CFD-based Eulerian-Lagrangian computational model that enables the rational design and optimization of such systems. The model takes into account dominant magnetic and hydrodynamic forces on the beads as well as coupled bead-fluid interactions. Fluid flow (Navier-Stokes equations) and mass transfer (Fick's law) between the coflowing fluids are solved numerically, while the magnetic force on the beads is predicted using analytical methods. The model is demonstrated via application to a prototype device and used to predict key performance metrics; degree of bead separation, flow patterns, and mass transfer, i.e. blood diffusion to the buffer phase. The impact of different process variables and parameters - flow rates, bead and magnet dimensions and fluid viscosities - on both bead recovery and blood loss or dilution is quantified for the first time. The performance of the prototype device is characterized using fluorescence microscopy and the experimental results are found to match theoretical predictions within an absolute error of 15%. While the model is demonstrated here for analysis of a detoxification device, it can be readily adapted to a broad range of magnetically-enabled microfluidic applications, e.g. bioseparation, sorting and sensing.


Pseudomonas piscium sp. nov., Pseudomonas pisciculturae sp. nov., Pseudomonas mucoides sp. nov. and Pseudomonas neuropathica sp. nov. isolated from rainbow trout.
Six Gram negative, motile bacteria were isolated from rainbow trout (Oncorhynchus mykiss). The 16S rRNA sequence similarity values grouped them in the Pseudomonas mandelii (strains P49, P50T, 154aT and P154b), Pseudomonas fluorescens (strain P115T) and Pseudomonas koreensis (strain P155T) phylogenetic subgroups in the genus Pseudomonas. The DNA G+C content ranged from 58.5 to 60 mol%. The strains were characterized phenotypically using API 20NE and Biolog GENIII tests, and chemotaxonomically by their whole-cell MALDI-TOF MS protein profiles and fatty acid contents. Multi-locus sequence analysis with four housekeeping gene sequences (rpoD, rpoB, gyrB and 16S rRNA) together with genome comparisons by average nucleotide identity and genome-to-genome distance calculations were performed. Results showed that the similarity values of these strains to known species type strains were lower than the thresholds established for species in the genus Pseudomonas. Based on these data, we concluded that strains P49, P50T, P115T, P154aT, P154b and P155T belonged to four novel species. The names proposed are: Pseudomonas piscium sp. nov. for strains P49 and P50T with P50T (=CECT 30175T=CCUG 74871T) as the type strain; Pseudomonas pisciculturae sp. nov. for strain P115T (CECT 30173T=CCUG 74873T); Pseudomonas mucoides sp. nov. for strains P154aT and P154b with P154aT (=CECT 30177T=CCUG 74874T) as the type strain; and Pseudomonas neuropathica sp. nov. for strain P155T (=CECT 30178T=CCUG 74875T).


Limitations of theophylline in the treatment of apnea of prematurity.
Theophylline is commonly used to treat apnea of prematurity. To determine the effectiveness of theophylline with respect to gestational and postnatal age, we conducted a controlled study in 43 premature infants with idiopathic apnea. Three of the 21 treated and eight of the 22 control infants developed respiratory failure. Eight of the 11 infants with respiratory failure had gestational ages of less than 31 weeks and had more than four apneic episodes during the first day of life. In the treated infants without respiratory failure, the number of apneic episodes by 24-hour intervals declined six days earlier than in the control infants. Apnea disappeared, however, at a similar time in both groups. Three treated infants and three control infants had apneic episodes persisting beyond the neonatal period. Patients treated with theophylline did not develop respiratory failure as often as control infants did. However, despite a reduction of apneic episodes, theophylline did not shorten the course of apnea of prematurity.


Artificial Pancreas: Clinical Study in Latin America Without Premeal Insulin Boluses.
Emerging therapies such as closed-loop (CL) glucose control, also known as artificial pancreas (AP) systems, have shown significant improvement in type 1 diabetes mellitus (T1DM) management. However, demanding patient intervention is still required, particularly at meal times. To reduce treatment burden, the automatic regulation of glucose (ARG) algorithm mitigates postprandial glucose excursions without feedforward insulin boluses. This work assesses feasibility of this new strategy in a clinical trial. A 36-hour pilot study was performed on five T1DM subjects to validate the ARG algorithm. Subjects wore a subcutaneous continuous glucose monitor (CGM) and an insulin pump. Insulin delivery was solely commanded by the ARG algorithm, without premeal insulin boluses. This was the first clinical trial in Latin America to validate an AP controller. For the total 36-hour period, results were as follows: average time of CGM readings in range 70-250 mg/dl: 88.6%, in range 70-180 mg/dl: 74.7%, <70 mg/dl: 5.8%, and <50 mg/dl: 0.8%. Results improved analyzing the final 15-hour period of this trial. In that case, the time spent in range was 70-250 mg/dl: 94.7%, in range 70-180 mg/dl: 82.6%, <70 mg/dl: 4.1%, and <50 mg/dl: 0.2%. During the last night the time spent in range was 70-250 mg/dl: 95%, in range 70-180 mg/dl: 87.7%, <70 mg/dl: 5.0%, and <50 mg/dl: 0.0%. No severe hypoglycemia occurred. No serious adverse events were reported. The ARG algorithm was successfully validated in a pilot clinical trial, encouraging further tests with a larger number of patients and in outpatient settings.


Circadian cortisol secretion in adolescent girls with conduct disorder.
Severe antisocial behavior in girls, best exemplified by conduct disorder (CD), is a serious clinical and public health problem. Treatment is difficult, particularly in girls with comorbid internalizing disorders. Identifying biological correlates may help to develop new treatments or diagnostic, prognostic, or treatment response biomarkers. Based on our earlier work and research from others occurring primarily in boys with severe antisocial behavior, it is possible that abnormalities in the hypothalamic pituitary adrenal (HPA) axis circadian cortisol cycle may be associated with female CD. Additionally, research suggests that the presence of comorbid internalizing disorders may be related to differences in cortisol secretion, compared to subjects who only have CD. Our study aimed: 1) to compare the circadian cortisol cycle in 98 girls with CD, 15-16 years of age to 47 girls without any psychiatric disorder (ND) and 2) to compare the cycle in girls with CD and comorbid internalizing disorders (CD + INT) to those without such comorbidity (CD Only). Salivary cortisol was collected over 24 h during weekdays at scheduled times, with protocol adherence measures in place. Unstructured covariance pattern modeling, controlling for effects of age, social class, IQ, and awakening time was used to analyze cortisol data. CD was associated with overall lower cortisol secretion (p = 0.03), but this difference was due to a lower volume of cortisol secreted 30 min after awakening (area under the curve with respect to ground, p = 0.01). Circadian cortisol secretion was no different in the CD+INT group compared to the CD Only group (p = 0.52). Our findings need to be replicated using current consensus guidelines for the assessment of the CAR. We also suggest two new avenues of research in this field.


Removal of Trihalomethanes by Dual Filtering Media (GAC-Sand) at El-Manshia Water Purification Plant.
Prechlorination is used as an initial step in water purification for public supply. One of the drawbacks of the prechlorination is the reaction between natural organic matters with chlorine forming trihalmethanes. This study aims at evaluating the performance of granular activated carbon (GAC) with sand as a dual filtering media with different depths on removal of trihalomethanes (THMs) for improving water quality. The Czeck sand filter at El-Manshia Water Purification Plant was chosen in this study in order to improve its water quality. The pilot filter was designed to work as mono medium sand filter and dual GAC-Sand media. The depths of GAC were 5 cm, 10 cm, 15 cm, 20 cm, 30 cm, and 40 cm over 115 cm, 110 cm, 105 cm, 100 cm, 90 cm, and 80 cm of sand, respectively. The six filter depths of GAC in the dual filter were studied to choose the optimum depth of GAC to improve water quality especially for THMs removal and comparing with mono-sand media and with Czeck filter. The results showed that the GAC-Sand dual media filter of 30 cm depth of GAC and 90 cm sand was the best depth for improving water quality where it was efficient in adsorbing mostly the total trihalomethanes in which its percentage of removal was 87%. The filtered water turbidity had an average of 0.3 NTU and its percentage of removal was 90%, algae removal was 95%, but it had a poor effect on bacteria removal with 27% removal due to adsorption of residual chlorine by GAC. The study recommended replacing mono media by dual media filter to improve water quality where the GAC was efficient to remove trihalomethanes in which the relative concentration (C/Co) was 0.16. The benefit cost calculated on 30 cm depth of GAC is equal to 0.04 piaster/m(3). In addition, it resulted in longer filter run of 54 hrs compared to average filter run of 24 hr for Czech filters, as well as increased water productivity where unit filter run volume was 324 m(3)/m(2) instead of 144 m(3)/m(2) for Czech mono media.


Improving Visualization Interpretation Using Counterfactuals.
Complex, high-dimensional data is used in a wide range of domains to explore problems and make decisions. Analysis of high-dimensional data, however, is vulnerable to the hidden influence of confounding variables, especially as users apply ad hoc filtering operations to visualize only specific subsets of an entire dataset. Thus, visual data-driven analysis can mislead users and encourage mistaken assumptions about causality or the strength of relationships between features. This work introduces a novel visual approach designed to reveal the presence of confounding variables via counterfactual possibilities during visual data analysis. It is implemented in CoFact, an interactive visualization prototype that determines and visualizes counterfactual subsets to better support user exploration of feature relationships. Using publicly available datasets, we conducted a controlled user study to demonstrate the effectiveness of our approach; the results indicate that users exposed to counterfactual visualizations formed more careful judgments about feature-to-outcome relationships.


[Inadvertent child health neglect by preference of homeopathy to conventional medicine].
The effectiveness of alternative medicine, especially homeopathy, has not been proven by prospective, double-blind, controlled studies. However, homeopathy has been gaining in popularity in Israel. 4 children of religious parents are presented, who were treated by homeopathic medical practitioners for cervical streptococcal lymphadenitis, acute lymphatic leukemia, bacterial pneumonia and atopic dermatitis, respectively. The parents of the latter 3 children rejected the correct medical diagnoses, as well as any medical treatment offered by other medical practitioners. All of the children had severe, unusual, but preventable complications. "Alternative" medicine, especially homeopathic practice, is dealt with from medical, social and comparative law aspects. The 4 cases are considered examples of a special kind of child neglect, inadvertent child health neglect because of false beliefs or lack of knowledge. The government is called upon to prevent this kind of child neglect by education of the public and the enactment of appropriate laws and their enforcement. This should be done even if the right to choose one's health treatment system and practitioner is preferred to the right not to be offered ineffective treatment.


Multivessel coronary revascularization in patients with and without diabetes mellitus: 3-year follow-up of the ARTS-II (Arterial Revascularization Therapies Study-Part II) trial.
The purpose of this study was to assess the 3-year outcome of coronary artery bypass graft surgery (CABG) and percutaneous coronary intervention (PCI) using sirolimus-eluting stents (SES) in patients who had multivessel coronary artery disease with and without diabetes mellitus. The optimal method of revascularization in diabetic patients remains in dispute. The ARTS-II (Arterial Revascularization Therapies Study-Part II) trial is a single-arm study (n = 607) that included 159 diabetic patients treated with SES whose 3-year clinical outcome was compared with that of the historical diabetic and nondiabetic arms of the randomized ARTS-I trial (n = 1,205, including 96 diabetic patients in the CABG arm and 112 in the PCI arm). At 3 years, among nondiabetic patients, the incidence of the primary composite of death, CVA, myocardial infarction (MI), and repeat revascularization (major adverse cardiac and cerebrovascular events [MACCE]), was significantly lower in ARTS-II than in ARTS-I PCI (adjusted odds ratio [OR]: 0.41; 95% confidence interval [CI]: 0.26 to 0.64) and similar to ARTS-I CABG. The ARTS-II patients were at significantly lower risk for death, CVA, and MI as compared with both the ARTS-I PCI (adjusted OR: 0.55; 95% CI: 0.34 to 0.91) and ARTS-I CABG patients (adjusted OR: 0.56; 95% CI: 0.35 to 0.92). Among diabetic patients, the incidence of MACCE in ARTS-II was similar to that of both PCI and CABG in ARTS-I. Conversely, the incidence of death, CVA, and MI was significantly lower in ARTS-II than in ARTS-I PCI (adjusted OR: 0.67; 95% CI: 0.27 to 1.65) and was similar to that of ARTS-I CABG. At 3 years, PCI using SES for patients with multivessel coronary artery disease seems to be safer and more efficacious than PCI using bare-metal stents, irrespective of the diabetic status of the patient. Hence, PCI using SES appears to be a valuable alternative to CABG for both diabetic and nondiabetic patients.


Clinicopathological predictors of positive 68Ga-PSMA-11 PET/CT in PSA-only recurrence of localized prostate cancer following definitive therapy.
To demonstrate the effect of clinicopathological factors on 68Ga-PSMA-11 PET/CT positivity at the time of biochemical recurrence (BCR) of localized prostate cancer (PCa) following definitive therapy. We retrospectively reviewed our institutional database for PCa patients who had BCR and subsequently underwent 68Ga-PSMA-11 PET/CT between April 2014 and February 2018. A total of 51 patients who were metastasis-free before PSMA imaging and previously treated with definitive therapy (radical prostatectomy or external beam radiotherapy) for localized disease (pT1c-T3b pN0-1 cM0) were included. 37 out of 51 patients (72.5%) had positive 68Ga-PSMA-11 PET/CT scans. Age at diagnosis, Gleason score (GS), D'Amico risk status of PCa, initial PSA level before treatment and PSA doubling time were not associated with PSMA positivity. Pre-scan PSA levels of > 0.2 ng/ml and PSA velocity of ≥ 1 ng/ml/year were significantly associated with increased PSMA positivity, whereas history of androgen deprivation therapy showed a trend towards significance. The optimal cutoffs for distinguishing between positive and negative scans were ≥ 0.71 ng/ml for pre-scan PSA and ≥ 1.22 ng/ml/yr for PSA velocity. In multivariable analysis, log pre-scan PSA and pre-scan PSA level > 0.2 ng/ml remained significant predictors for PSMA positivity, whereas the association of PSA velocity and of ADT was lost. In BCR of localized PCa following definitive therapy, pre-scan PSA was strongly associated with positive 68Ga-PSMA-11 scan, even at PSA levels ranging from 0.2 to 1.0 ng/ml. Therefore, clinical and pathological predictors of positive 68Ga-PSMA-11 PET/CT in PSA-only recurrence of localized prostate cancer need to be further elucidated.


The beta-lactamase of Enterobacter cloacae P99. Chemical properties, N-terminal sequence and interaction with 6 beta-halogenopenicillanates.
The beta-lactamase of Enterobacter cloacae P99 consists of one polypeptide chain of Mr 39000 devoid of disulphide bridges and free thiol groups. It contains an unusually high proportion of tyrosine and tryptophan. The N-terminal sequence exhibits overlaps with the tryptic peptide obtained after labelling the active site with 6 beta-iodopenicillanate. The active-site serine residue is at position 64. The homology with the chromosomal beta-lactamase of Escherichia coli K 12 (ampC gene) is lower within the 25 residues of the N-terminal portion than around the active-site serine residue. The P99 beta-lactamase is inactivated by 6 beta-bromo- and 6 beta-iodo-penicillanate, with a second-order rate constant of 110-140M-1 X s-1 at 30 degrees C and pH 7.0, a value that is much lower than that observed with class-A beta-lactamases.


Computed tomography-estimated specific gravity at hospital admission predicts 6-month outcome in mild-to-moderate traumatic brain injury patients admitted to the intensive care unit.
It is clear that patients with a severe traumatic brain injury (TBI) develop secondary, potentially lethal neurological deterioration. However, it is difficult to predict which patients with mild-to-moderate TBI (MM-TBI), even after intensive care unit (ICU) admission, will experience poor outcome at 6 months. Standard computed tomography (CT) imaging scans provide information that can be used to estimate specific gravity (eSG). We have previously demonstrated that higher eSG measurements in the standard CT reading were associated with poor outcomes after severe TBI. The aim of this study was to determine whether eSG of the intracranial content predicts 6-month outcome in MM-TBI. We analyzed admission clinical and CT scan data (including eSG) of 66 patients with MM-TBI subsequently admitted to our neurosurgical ICU. Primary outcome was defined as a Glasgow Outcome Scale score of 1 to 3 after 6 months. Discriminating power (area under the receiver operating characteristic curve [ROC-AUC], 95% confidence interval) of eSG to predict 6-month poor outcome was calculated. The correlation of eSG with the main ICU characteristics was then compared. Univariate and stepwise multivariate analyses showed an independent association between eSG and 6-month poor outcome (P = 0.001). ROC-AUC of eSG for the prediction of 6-month outcomes was 0.87 (confidence interval: 0.77-0.96). Admission eSG values were correlated with the main ICU characteristics, specifically 14-day mortality (P = 0.004), length of mechanical ventilation (P = 0.01), length of ICU stay (P = 0.045), and ICU procedures such as intracranial pressure monitoring (P < 0.001). In this MM-TBI cohort admitted to the ICU, eSG of routine CT scans was correlated with mortality, ICU severity, and predicted 6-month poor outcome. An external validation with studies that include the spectrum of TBI severities is warranted to confirm our results.


Expandable Technology Improves Clinical and Radiographic Outcomes of Minimally Invasive Lateral Lumbar Interbody Fusion for Degenerative Disc Disease.
Static interbody spacers are standard of care for minimally invasive lateral lumbar interbody fusion (MIS LLIF). However, placement of large static interbody spacers typically requires multiple trialing, endplate preparation, and forceful impaction. A lateral expandable interbody spacer with adjustable lordosis can be inserted at a reduced height, to optimize the endplate-to-endplate fit. This study describes radiographic and clinical outcomes in patients treated using lateral titanium expandable interbody spacers with adjustable lordosis using MIS LLIF. This is a single-surgeon, retrospective, institutional review board-exempt chart review of 24 consecutive patients who underwent MIS LLIF at 1-2 contiguous level(s) using expandable spacers with adjustable lordosis. Radiographic and clinical functional outcomes were collected and compared at preoperative and postoperative time points up to 24 months. Parametric and nonparametric tests were used when appropriate. Statistical results were significant if P < .05. Twenty-four consecutive patients were evaluated with an average age of 57.8 ± 12.6 years; 45.8% were female. Visual analog scale for back pain improved by 7.3 ± 1.0 points, whereas Oswestry Disability Index scores improved by a mean of 67.5 ± 11.3 points at 24 months (P < .001). Lumbar lordosis improved by a mean of 6.3 ± 10.1° at 24 months (P < .001). There were 29 spinal levels, with 41.4% at L4-5 and 34.5% at L3-4. Anterior, middle, and posterior disc height significantly increased at 24 months by means of 4.5 ± 2.9 mm, 4.0 ± 2.8 mm, and 2.6 ± 1.9 mm, respectively (P < .001). Neuroforaminal height significantly improved by 3.3 ± 3.9 mm at 24 months (P < .001). Segmental lordosis improved by 3.6 ± 3.0° at 24 months. This study showed significant positive clinical and radiographic outcomes for patients who underwent MIS LLIF using expandable interbody spacers with adjustable lordosis. Correction of sagittal alignment was achieved and maintained up to 2-year follow-up. The use of expandable spacers with adjustable lordosis was shown to be safe and effective in this cohort. 3.


[Familial study of patients with hereditary hemorrhagic telangiectasia].
Rendu-Osler-Weber's disease or Hereditary Hemorrhagic Telangiectasia (HTT) is an autosomal dominant hereditary clinical entity characterized by the presence of telangiectasias on skin, mucous membranes and internal organs. The incidence of hepatic, pulmonary or cerebral complications justifies an early diagnostic. A familiar study of patients previously diagnosed of HTT in our hospital was made, establishing a protocol for the identification of asymptomatic or pauci-symptomatic cases. Fourteen patients were studied: 6 with a previous diagnostic, and 8 familiar contacts. Penetrance was of 85%. Epistaxis (80%) and telangiectasias on skin were the most frequent clinical findings. The visceral lesions found were gastrointestinal (28%), hepatic (15%), urological (15%) and pulmonary (7%). All patients were included in an hepatitis B virus vaccination program. It was also made a morphological study with Werhoeff's staining of the elastic layer, that allowed to distinguish both arterial and venous alterations.


Naive and memory CD4+ T cell subsets can contribute to the generation of human Tfh cells.
CD4+ T follicular helper cells (Tfh) promote B cell maturation and antibody production in secondary lymphoid organs. By using an innovative culture system based on splenocyte stimulation, we studied the dynamics of naive and memory CD4+ T cells during the generation of a Tfh cell response. We found that both naive and memory CD4+ T cells can acquire phenotypic and functional features of Tfh cells. Moreover, we show here that the transition of memory as well as naive CD4+ T cells into the Tfh cell profile is supported by the expression of pro-Tfh genes, including transcription factors known to orchestrate Tfh cell development. Using this culture system, we provide pieces of evidence that HIV infection differentially alters these newly identified pathways of Tfh cell generation. Such diversity in pathways of Tfh cell generation offers a new framework for the understanding of Tfh cell responses in physiological and pathological contexts.


[New class of medicines for chronic hepatitis C].
Worldwide approximately 130-210 million people suffer from chronic hepatitis C. Adequate antiviral therapy reduces morbidity and mortality caused by chronic hepatitis C and prevents further spread of the hepatitis C-virus (HCV). The current standard treatment of chronic hepatitis C, consisting of the combination of pegylated interferon-α (peginterferon) and ribavirin, lasts 24-48 weeks, and is accompanied by significant side effects and has a suboptimal chance of success. Protease inhibitors, which have recently been registered, belong to a new class of medicines which directly affect the life cycle of HCV. Protease inhibitors, in combination with peginterferon and ribavirin, provide almost double the chance of curing in patients with HCV genotype 1. Treatment duration can be shortened in a considerable proportion of these patients. Since treatment with protease inhibitors can lead to resistant virus strains and this therapy leads to additional side effects, the complexity of treatment will increase.


Extracellular matrix changes PC12 cell shape and processing of newly synthesized dopamine.
PC12 cells on extracellular matrix (ECM) or plastic were incubated with 3H-tyrosine (3H-TY) in the presence and absence of serum or cold tyrosine. 3H-Dopamine (3H-DA) was determined in medium and cells from 1 to 48 h later with Dowex cation exchange chromatography. In serum-free and tyrosine-free medium, PC12 cells on ECM released significantly more 3H-DA, whereas cells on plastic had a significantly higher cellular content of 3H-DA, but total 3H-DA (medium plus cells) was equal in ECM and plastic cultures. When 3H-TY was added to tyrosine-containing medium, there was a significant decrease in the levels of 3H-DA detected and the differences between ECM and plastic cultures were attenuated, but the patterns of secretion and storage were similar to those observed with tyrosine-free medium and total synthesis did not decline at 48 h. Serum decreased the efficiency of the resin to retain 3H-DA from culture medium, attenuated the difference in dopamine release between ECM and plastic cultures, and contributed to variations in 3H-TY uptake. The morphometric relationship between the cell membrane and the internal compartment in PC12 cells of different shapes was also characterized. The perimeter length and area of the midsection of cells were determined with a modular system for quantitative digital analysis. The perimeter length of cells on ECM was significantly greater than cells on plastic, whereas the internal areas were similar. The ratio of perimeter length to area (P/A) for all cells on ECM was 30% higher than the P/A ratio for cells on plastic. The ratio of P/A for a subpopulation of very flat cells on ECM was 70% higher than the ratio for round cells on plastic. Immunocytochemistry for tyrosine hydroxylase revealed a more diffuse distribution of this enzyme in cells on ECM. These data suggest that there is an increase in the ratio of cell surface area to cell volume as PC12 cells spread on ECM which could facilitate secretory vesicle fusion with the cell membrane, and hence, exocytosis. Although there is a concomitant increase in the secretion of dopamine and a decrease in the storage of dopamine, the change in cell shape does not appear to immediately alter the synthesis of dopamine.


Community-Based Participatory Research to Adapt Health Measures for Use by People With Developmental Disabilities.
People with developmental disabilities (DD) are often not included as participants in research owing to a variety of ethical and practical challenges. One major challenge is that traditional measurement instruments may not be accessible to people with DD. Participatory research approaches promise to increase the participation of marginalized communities in research, but few partnerships have successfully used such approaches to conduct quantitative studies people with DD. To use a community-based participatory research (CBPR) approach to create an accessible, computer-assisted survey about violence and health in people with DD, and to psychometrically test adapted health instruments. Our academic-community partnership, composed of academic researchers, people with DD, and supporters, collaboratively selected and modified data collection instruments, conducted cognitive interviews and pilot tests, and then administered the full survey to 350 people with DD. Although team members sometimes had opposing accommodation needs and adaptation recommendations, academic and community partners were able to work together successfully to adapt instruments to be accessible to participants with a wide range of DD. Results suggest the adapted health instruments had strong content validity and all but one had good to excellent internal consistency reliability (alpha, 0.81-0.94). The majority of participants (75%) responded that all or most of the questions were easy to understand. Researchers should consider using participatory approaches to adapting instruments so people with DD can be validly included in research.


Membranous expression of podocalyxin-like protein is an independent factor of poor prognosis in urothelial bladder cancer.
Membranous expression of the anti-adhesive glycoprotein podocalyxin-like (PODXL) has previously been found to correlate with poor prognosis in several major cancer forms. Here we examined the prognostic impact of PODXL expression in urothelial bladder cancer. Immunohistochemical PODXL expression was examined in tissue microarrays with tumours from two independent cohorts of patients with urothelial bladder cancer: n=100 (Cohort I) and n=343 (Cohort II). The impact of PODXL expression on disease-specific survival (DSS; Cohort II), 5-year overall survival (OS; both cohorts) and 2-year progression-free survival (PFS; Cohort II) was assessed. Membranous PODXL expression was significantly associated with more advanced tumour (T) stage and high-grade tumours in both cohorts, and a significantly reduced 5-year OS (unadjusted HR=2.25 in Cohort I and 3.10 in Cohort II, adjusted HR=2.05 in Cohort I and 2.18 in Cohort II) and DSS (unadjusted HR=4.36, adjusted HR=2.70). In patients with Ta and T1 tumours, membranous PODXL expression was an independent predictor of a reduced 2-year PFS (unadjusted HR=6.19, adjusted HR=4.60) and DSS (unadjusted HR=8.34, adjusted HR=7.16). Membranous PODXL expression is an independent risk factor for progressive disease and death in patients with urothelial bladder cancer.


Recent advances in the treatment of colonic diverticular disease and prevention of acute diverticulitis.
The incidence of diverticulosis and diverticular disease of the colon is increasing worldwide. Although the majority of patients remains asymptomatic long-life, the prevalence of diverticular disease of the colon, including acute diverticulitis, is substantial and is becoming a significant burden on National Health Systems in terms of direct and indirect costs. Focus is now being drawn on identifying the correct therapeutic approach by testing various treatments. Fiber, non-absorbable antibiotics and probiotics seem to be effective in treating symptomatic and uncomplicated patients, and 5-aminosalicylic acid might help prevent acute diverticulitis. Unfortunately, robust evidence on the effectiveness of a medical strategy to prevent acute diverticulitis recurrence is still lacking. We herein provide a concise review on the effectiveness and future perspectives of these treatments.


Stable pollination service in a generalist high Arctic community despite the warming climate.
Insects provide key pollination services in most terrestrial biomes, but this service depends on a multistep interaction between insect and plant. An insect needs to visit a flower, receive pollen from the anthers, move to another conspecific flower, and finally deposit the pollen on a receptive stigma. Each of these steps may be affected by climate change, and focusing on only one of them (e.g., flower visitation) may miss important signals of change in service provision. In this study, we combine data on visitation, pollen transport, and single-visit pollen deposition to estimate functional outcomes in the high Arctic plant-pollinator network of Zackenberg, Northeast Greenland, a model system for global warming-associated impacts in pollination services. Over two decades of rapid climate warming, we sampled the network repeatedly: in 1996, 1997, 2010, 2011, and 2016. Although the flowering plant and insect communities and their interactions varied substantially between years, as expected based on highly variable Arctic weather, there was no detectable directional change in either the structure of flower-visitor networks or estimated pollen deposition. For flower-visitor networks compiled over a single week, species phenologies caused major within-year variation in network structure despite consistency across years. Weekly networks for the middle of the flowering season emerged as especially important because most pollination service can be expected to be provided by these large, highly nested networks. Our findings suggest that pollination ecosystem service in the high Arctic is remarkably resilient. This resilience may reflect the plasticity of Arctic biota as an adaptation to extreme and unpredictable weather. However, most pollination service was contributed by relatively few fly taxa (Diptera: Spilogona sanctipauli and Drymeia segnis [Muscidae] and species of Rhamphomyia [Empididae]). If these key pollinators are negatively affected by climate change, network structure and the pollination service that depends on it would be seriously compromised.


Dominance of orientation over frequency in the perception of 3-D slant and shape.
In images of textured three-dimensional surfaces, pattern changes can be characterized as changes in orientation and spatial frequency, features for which neurons in primary visual cortex are classically selective. Previously, we have demonstrated that correct 3-D shape perception is contingent on the visibility of orientation flows that run parallel to the surface curvature. We sought to determine the relative contributions of orientation modulations (OMs) and frequency modulations (FMs) for the detection of slant and shape from 3-D surfaces. Results show that 1) when OM and FM indicate inconsistent degrees of surface slant or curvature, observer responses were consistent with the slant or curvature specified by OM even if the FM indicated a slant or curvature in the opposite direction to the same degree. 2) For slanted surfaces, OM information dictates slant perception at both shallow and steep slants while FM information is effective only for steep slants. Together these results point to a dominant role of OM information in the perception of 3-D slant and shape.


Contrasting Hydraulic Architectures of Scots Pine and Sessile Oak at Their Southernmost Distribution Limits.
Many temperate European tree species have their southernmost distribution limits in the Mediterranean Basin. The projected climatic conditions, particularly an increase in dryness, might induce an altitudinal and latitudinal retreat at their southernmost distribution limit. Therefore, characterizing the morphological and physiological variability of temperate tree species under dry conditions is essential to understand species' responses to expected climate change. In this study, we compared branch-level hydraulic traits of four Scots pine and four sessile oak natural stands located at the western and central Mediterranean Basin to assess their adjustment to water limiting conditions. Hydraulic traits such as xylem- and leaf-specific maximum hydraulic conductivity (KS-MAX and KL-MAX), leaf-to-xylem area ratio (AL:AX) and functional xylem fraction (FX) were measured in July 2015 during a long and exceptionally dry summer. Additionally, xylem-specific native hydraulic conductivity (KS-N) and native percentage of loss of hydraulic conductivity (PLC) were measured for Scots pine. Interspecific differences in these hydraulic traits as well as intraspecific variability between sites were assessed. The influence of annual, summer and growing season site climatic aridity (P/PET) on intraspecific variability was investigated. Sessile oak displayed higher values of KS-MAX, KL-MAX, AL:AX but a smaller percentage of FX than Scots pines. Scots pine did not vary in any of the measured hydraulic traits across the sites, and PLC values were low for all sites, even during one of the warmest summers in the region. In contrast, sessile oak showed significant differences in KS-MAX, KL-MAX, and FX across sites, which were significantly related to site aridity. The striking similarity in the hydraulic traits across Scots pine sites suggests that no adjustment in hydraulic architecture was needed, likely as a consequence of a drought-avoidance strategy. In contrast, sessile oak displayed adjustments in the hydraulic architecture along an aridity gradient, pointing to a drought-tolerance strategy.


Orthogonal test design for optimization of the extraction of flavonid from the Fructus Gardeniae.
It is imperative to provide some consistent experimental results for the extraction of flavonid from Fructus Gardeniae. The key extraction parameters that influenced the yield of flavonid from Fructus Gardeniae were optimized by employing an orthogonal experiment [L(9)(3)(4)], including the ratio of buffer solution (Na(2)B(4)O(7)·10H(2)O) to raw material, concentration of Fructus Gardeniae in extracting solution, extraction time and pH of buffer solution. An UV/Vis detector was used to perform the qualitative and quantitative analyses of the extracted flavonid with the using of the standard sample. The maximum extraction yield of the crude extract was 5.0533 (mg/g) after 20 min when the mass ratio of Na(2)B(4)O(7)·10H(2)O to raw material was 0.4%, the concentration of Fructus Gardeniae in the extraction solution was 1/12 (g/mL), and pH of buffer solution was 4.5. The positive reactions to the Molish and HCl-Mg tests suggested that the extracted compound was flavonoid, and FTIR measurements also identified the presence of flavonoid in the extracts. This work is expected to provide a basis for further research, development, and utilization of Fructus gardenia in flavonid extraction.


Mortalin-based cytoplasmic sequestration of p53 in a nonmammalian cancer model.
In nature the soft shell clam Mya arenaria develops a fatal neoplasm that shares molecular similarity with an unrelated group of human cancers. In leukemic clam hemocytes, wild-type p53 and mortalin proteins co-localize in the cytoplasm. A similar phenotype, characterized by cytoplasmic sequestration of wild-type p53 protein, has been observed in several human cancers (undifferentiated neuroblastoma, retinoblastoma, colorectal and hepatocellular carcinomas, and glioblastoma). In some of these cancers p53 is tethered in the cytoplasm by mortalin when the latter protein is overexpressed. Using co-immunoprecipitation we have demonstrated that mortalin and p53 proteins are complexed in the cytoplasm of leukemic clam hemocytes (and not in normal hemocytes). In addition, treatment of leukemic clam hemocytes with MKT-077, a cationic inhibitor of mortalin, disrupts the interaction of mortalin and p53 proteins, resulting in translocation of some p53 to the nucleus. Based on these data, we introduce leukemic clam hemocytes as novel and easily accessible, in vivo and in vitro models for human cancers displaying a similar mortalin-based phenotype. Treatment of these models with novel chemotherapeutics may help reveal the molecular mechanism(s) involved in inactivating p53 by this form of cytoplasmic sequestration.


[Cross resistance in tumor cells and staphylococci with respect to anthracycline-group antineoplastic antibiotics].
The variants of the tumor cells of Fisher lymphadenosis, strain L-5178 and Staph. aureus resistant to rubomycin simultaneously became partially less sensitive to adriamycin. Sensitivity to karminomycin in the rubomycin resistant strains did not practically change as compared to the sensitivity of the initial strains. Sensitivity to adriamycin and rubomycin in Staph. aureus decreased 67 and 4 times respectively after 7 passages on media with increasing concentrations of adriamycin, while sensitivity to karminomycin decreased only 1.5 times, i.e. remained practically unchanged. After 22 passages of Staph. aureus to karminomycinrubomycin and adriamycin decreased 16, 67 and 33 times respectively. The results of the study may be explained by differences in the changes of the cell membrane permeability due to the drug effect.


[125 reinterventions for recurrent popliteal varicose veins after excision of the short saphenous vein. Anatomical and physiological hypotheses of the mechanism of recurrence].
Recurrence of popliteal varicose veins has long been attributed to insufficient excision of an incompetent short saphenous vein. Indeed, recurrence is still frequent after surgery of the short saphenous. In order to assess the risk of insufficient excision and study the mechanism of recurrence, we reviewed 125 popliteal procedures for recurrence after excision of an incompetent short saphenous vein. Among this series, 48 were personal procedures and among these, 43 had been performed without preoperative ultrasound explorations prior to 1991. Recurrences were classified into five categories (table I) according to the anatomic presentation at reoperation. Type 1) 17 patients had an intact short saphenous (13.6%) with either an inadequate incision or a simple recanalized suture. Type 2) 53 patients had a long stump (42.4%) with new superficial varicose communications. Type 3) 4 patients had a small residual and incompetent short saphenous trunk (3.2%), 20 patients had both a long stump and an incompetent residual trunk (16%). Type 4) 29 patients had incompetent popliteal perforating veins (23.2%). Type 5) 2 patients developed recurrence on a new varicose communication which followed the posterior nerves of the thigh. Among our personal series, 43 reoperations for recurrence were performed without preoperative duplex Doppler exploration. Among them, excision was insufficient in 38 (table II). The 5 patients reoperated after preoperative ultrasound exploration had a popliteal perforating vein. Recurrence with a popliteal perforator was significantly more frequent in men than in women (table III). The delay to reoperation for popliteal recurrence after surgery of the short saphenous (50% at 6 years) was significantly shorter than the delay to reoperation for inguinal recurrence after surgery of the greater saphenous (50% at 12 years) (table V). Insufficient excision was observed in 75.2% of the cases, but 23.2% of the recurrences were due to the development of an incompetent popliteal perforating veins. These perforating veins were residual stumps of the short saphenous with complex pathways, unrecognized perforating veins associated with the short saphenous at the first operation, or a new popliteal incompetence. Ces perforantes de la fosse poplitée pourraient être des moignons résiduels de petites saphènes à trajet complexe, des perforantes méconnues associées à la petite saphène developing in several perforating vessels described in the popliteal fossa. The lack of preoperative ultrasound data made it difficult to interpret these recurrences. The development of perforating veins in the popliteal fossa is a type of recurrence which is probably the expression of particular hemodynamic phenomena in the popliteal venous circulation. These phenomena probably involve the flexion of the popliteal vein, the contraction of the calf muscles, and also popliteal valvular incompetence frequently demonstrated in patients who develop short saphenous vein insufficiency.


First Report of Freesia sneak virus in Commercial Freesia hybrida Cultivars in Korea.
In order to investigate the virus infection rate of commercial freesia cultivars in early February 2013, 19 freesia cultivars showing necrotic purple speckles or streaks on leaves, purple streaks parallel to the midrib, and necrotic speckles on leaves were collected from three different regions (Suwon and Icheon in Gyeonggi Province and Jeonju in North Jeonla Province) and used for virus detection. Nucleic acid extracts were analyzed for detection of major freesia-infecting viruses including Freesia sneak virus (FreSV) by reverse transcription (RT)-PCR with specific primer pairs. The FreSV CP gene was amplified using primer pair FreSV-F (5'-TTAGATAGTGAATCCATAAGCTGC-3') and FreSV-R (5'-ATGTCTGGAAAATACTCCGTCCAA-3'). The approximately 1.3-kb fragment of the FreSV amplified product was cloned and sequenced (GenBank Accession No. KC771891 to 98). The nucleotide sequences of CP gene of FreSV korean isolates showed 99.2 to 99.8% similarity to other FreSV isolates DQ885455, FJ807730, and GU071089, which are registered in GenBank. FreSV was detected from 71.7% of 138 plants tested while the infection rate of Freesia mosaic virus (FreMV) was 34.8%. Neither Bean yellow mosaic virus (BYMV) nor Tobacco rattle virus (TRV) were detected from any plants tested in this study. In certain cultivars, such as 'Bluebau' (II) and 'Pretty women,' most plants planted in the field showed purple streak symptoms on the leaves. In conclusion, FreSV was detected from some symptomatic freesia cultivars showing purple streak or speckles on leaves with or without necrotic spots and necrotic speckles on leaves. FreSV is currently widespread in Korea and some freesia plants were mixed infected with FreMV. FreSV has been occurring in the Netherlands for over 40 years (2). It is a plant virus in the family Ophioviridae and Ophiovirus genus. Once it occurs in freesia plantation fields, eradication is almost impossible because FreSV is transmitted by zoospores of Olpidium brassicae, which is a soilborne root-infecting fungus (3). Resting spores of O. brassicae can remain dormant in the soil and can be infective for 20 years (1). To produce virus-free freesia plants, growers should consider whether or not their fields are contaminated with O. brassicae carrying FreSV. To our knowledge, this is the first report of FreSV in freesia plants in Korea. References: (1) R. N. Campbell. Can. J. Bot. 63:2288, 1985. (2) Y. Koot et al. Tijdschrift over Plantenziekten 60:157, 1954. (3) H. J. M. van Dorst. Neth. J. Plant Pathol. 81:45, 1975.