Speciation of zinc in fish feed by size exclusion chromatography coupled to inductively coupled plasma mass spectrometry - using fractional factorial design for method optimisation and mild extraction conditions.
Zinc (Zn) is an element essential to all living organisms and it has an important role as a cofactor of several enzymes. In fish, Zn deficiency has been associated with impaired growth, cataracts, skeletal abnormalities and reduced activity of various Zn metalloenzymes. Fish meal and fish oil traditionally used in salmon feed preparation are being replaced by plant-based ingredients. Zinc additives are supplemented to salmon feed to ensure adequate Zn levels, promoting good health and welfare in Atlantic salmon (Salmo salar). The main objective of the present study was to evaluate Zn species found in an Atlantic salmon feed. This work describes a Zn extraction method that was optimized using a fractional factorial design (FFD), whereby the effect of six factors could be studied by performing only eight experiments. The effects of the type of extraction solution and its molar concentration, pH, presence of sodium dodecyl sulphate, temperature and extraction time on Zn extraction were investigated. Mild extraction conditions were chosen in order to keep the Zn species intact. Total Zn (soluble fractions and non-soluble fractions) was determined by inductively coupled plasma mass spectrometry (ICP-MS). The highest Zn recovery was obtained using 100 mM Tris-HCl, pH 8.5 at a temperature of 4 °C for 24 h where the total Zn in soluble fraction and non-soluble fraction was 9.9 ± 0.2% and 98 ± 6%, respectively. Zinc speciation analysis (on the soluble fractions) was further conducted by size exclusion inductively coupled plasma mass spectroscopy (SEC-ICP-MS). The SEC-ICP-MS method provided qualitative and semi-quantitative information regarding Zn species present in the soluble fractions of the feed. Four Zn-containing peaks were found, each with different molecular weights: Peak 1 (high molecular weight - ≥600 kDa), peak 2 and peak 3 (medium molecular weight - 32 to 17 kDa) were the least abundant (1-6%), while peak 4 (low molecular weight - 17 to 1.36 kDa) was the most abundant (84-95%).


From words to complete phrases: insight into single-cell isoforms using short and long reads.
The profiling of gene expression patterns to glean biological insights from single cells has become commonplace over the last few years. However, this approach overlooks the transcript contents that can differ between individual cells and cell populations. In this review, we describe early work in the field of single-cell short-read sequencing as well as full-length isoforms from single cells. We then describe recent work in single-cell long-read sequencing wherein some transcript elements have been observed to work in tandem. Based on earlier work in bulk tissue, we motivate the study of combination patterns of other RNA variables. Given that we are still blind to some aspects of isoform biology, we suggest possible future avenues such as CRISPR screens which can further illuminate the function of RNA variables in distinct cell populations.


IntS6 and the Integrator phosphatase module tune the efficiency of select premature transcription termination events.
The metazoan-specific Integrator complex catalyzes 3' end processing of small nuclear RNAs (snRNAs) and premature termination that attenuates the transcription of many protein-coding genes. Integrator has RNA endonuclease and protein phosphatase activities, but it remains unclear if both are required for complex function. Here, we show IntS6 (Integrator subunit 6) over-expression blocks Integrator function at a subset of Drosophila protein-coding genes, although having no effect on snRNAs or attenuation of other loci. Over-expressed IntS6 titrates protein phosphatase 2A (PP2A) subunits, thereby only affecting gene loci where phosphatase activity is necessary for Integrator function. IntS6 functions analogous to a PP2A regulatory B subunit as over-expression of canonical B subunits, which do not bind Integrator, is also sufficient to inhibit Integrator activity. These results show that the phosphatase module is critical at only a subset of Integrator-regulated genes and point to PP2A recruitment as a tunable step that modulates transcription termination efficiency.


Vitamin D is associated with testosterone and hypogonadism in Chinese men: Results from a cross-sectional SPECT-China study.
To date, no study has explored the association between androgen levels and 25-hydroxyvitamin D (25(OH)D) levels in Chinese men. We aimed to investigate the relationship between 25(OH)D levels and total and free testosterone (T), sex hormone binding globulin (SHBG), estradiol, and hypogonadism in Chinese men. Our data, which were based on the population, were collected from 16 sites in East China. There were 2,854 men enrolled in the study, with a mean (SD) age of 53.0 (13.5) years. Hypogonadism was defined as total T <11.3 nmol/L or free T <22.56 pmol/L. The 25(OH)D, follicle-stimulating hormone, luteinizing hormone, total T, estradiol and SHBG were measured using chemiluminescence and free T by enzyme-linked immune-sorbent assay. The associations between 25(OH)D and reproductive hormones and hypogonadism were analyzed using linear regression and binary logistic regression analyses, respectively. A total of 713 (25.0 %) men had hypogonadism with significantly lower 25(OH)D levels but greater BMI and HOMA-IR. Using linear regression, after fully adjusting for age, residence area, economic status, smoking, BMI, HOMA-IR, diabetes and systolic pressure, 25(OH)D was associated with total T and estradiol (P < 0.05). In the logistic regression analyses, increased quartiles of 25(OH)D were associated with significantly decreased odds ratios of hypogonadism (P for trend <0.01). This association, which was considerably attenuated by BMI and HOMA-IR, persisted in the fully adjusted model (P for trend <0.01) in which for the lowest compared with the highest quartile of 25(OH)D, the odds ratio of hypogonadism was 1.50 (95 % CI, 1.14, 1.97). A lower vitamin D level was associated with a higher prevalence of hypogonadism in Chinese men. This association might, in part, be explained by adiposity and insulin resistance and warrants additional investigation.


Gonadotropin responses following luteinizing hormone releasing hormone administration in normal subjects.
Synthetic luteinizing hormone releasing hormone (LHRH), 100 mug, was administered intravenously to 29 normal individuals. In male subjects, both baseline and peak LH response values showed minimal variations between subjects. The FSH baseline and response were more variable. In normal females the FSH and LH baseline values and response to LHRH infusion showed greater variation than that seen in male subjects, and this variation correlated with the phase of the menstrual cycle. In perimenopausal women with elevated baseline gonadotropins, the absolute LH peak values were comparable to those seen during the periovulatory period of the menstrual cycle, while FSH baseline and peak values were unmistakably higher than those observed in the normally menstruating women. Three women under gonadotropic suppression by three different drugs were investigated; megestrol suppressed the LHRH response more effectively than either diethylstilbestrol or an oral contraceptive. The LHRH stimulation test reflects only the ability of the pituitary cell to release gonadotropins. In a normal individual, however, some justifiable assumptions can be made regarding the immediate past synthesis and release versus accumulation of pituitary LH.


Problem- vs. lecture-based pharmacology teaching in a German medical school.
We have compared the effectiveness of problem-based learning (PBL) and classical lecture-based learning (LBL) in conveying medical facts in a general pharmacology class of third year medical students ( n=107). Three groups with a total of 31 students were randomly assigned to PBL. The PBL groups (9-12 students each) received ten 2-h sessions in which a clinical case was discussed and ten 2-h sessions in which areas of pharmacology not covered by the case discussions were presented in an LBL format (one group with all 31 students). The other students were assigned to groups of 14-15 students and received 20 2-h sessions in an LBL format. At the end of the semester all students received a questionnaire and participated in the same 30-question multiple-choice exam. The mean number of false answers was 7.6+/-4.0 and 9.7+/-4.7 in the PBL and LBL groups, respectively ( P<0.05 in a two-tailed t-test), and the percentage of failing students (>10 false answers) was 27% and 38%, respectively. Both groups were asked to rate their pharmacology class on a scale of 1 (lowest) to 10 (highest). In this questionnaire, PBL students by average rated generated interest in pharmacology, conveyed knowledge in pharmacology and understanding of medical questions approximately 1 point higher than LBL students. In an additional questionnaire given to the PBL students only, they reported to have prepared themselves 0.9+/-1.1 h for their lecture sessions but 3.0+/-1.4 h for their case discussions. The above findings were largely confirmed in the next semester when students were allowed to decide whether to participate in PBL or LBL classes. Moreover, PBL students did not score worse than LBL students in nation-wide pharmacology exams (523+/-76 vs. 500+/-91 points, P=0.09). We conclude that a switch from LBL to PBL teaching of pharmacology does not occur at the expense of factual knowledge transmission.


Nitrofurantoin-induced hepatotoxicity: a rare yet serious complication.
Nitrofurantoin is a commonly prescribed antibiotic for the treatment of recurrent uncomplicated urinary tract infections. Its importance has been emphasized by the current international clinical practice guidelines for the management of uncomplicated cystitis. Since its introduction into clinical practice, nitrofurantoin has been associated with various adverse effects, including hepatotoxicity. We searched the English-language literature using PubMed and SCOPUS for the period 1961 through the end of February 2013. Key search terms included "nitrofurantoin AND hepatotoxicity" as well as "nitrofurantoin AND hepatitis." When studies or case reports were found, we assessed articles cited in those publications. A broad spectrum of liver toxicity associated with nitrofurantoin use has been reported, ranging from acute hepatitis, granulomatous reaction, cholestasis, or autoimmune-mediated hepatitis to chronic active hepatitis that could lead to cirrhosis or death. The mechanism of hepatotoxicity is poorly understood, but it is believed to be the result of an immunologic process or a direct cytotoxic reaction. It has been postulated that prolonged exposure to nitrofurantoin, female sex, advanced age, and reduced renal function increase the risk of developing hepatotoxicity. For the management of severe cases, corticosteroids have been used along with nitrofurantoin discontinuation. Because of mixed results, the utility of corticosteroids has not been proven and should be used judiciously. Given the severity and seriousness of the adverse effect of hepatotoxicity, clinicians should weigh the risks and benefits of nitrofurantoin before initiating therapy, especially in long-term prophylaxis in high-risk patients. Clinicians also should be well versed in recognizing and managing liver injury associated with nitrofurantoin.


Boron neutron capture therapy for newly diagnosed glioblastoma multiforme: an assessment of clinical potential.
The purpose of this analysis was to assess the potential of BNCT, with L-boronophenylalanine (L-BPA), as first line radiotherapy for glioblastoma multiforme (GBM). The survival of patients with newly diagnosed GBM from a phase II BNCT study was compared with those from the two arms of a phase III study with conventional radiotherapy (RT) vs. RT plus concomitant and adjuvant medication with temozolomide (TMZ). A small subgroup, for which the methylation status of the O(6)-methylguanine-DNA methyltransferase (MGMT) DNA-repair gene was known, was also considered. The results indicated that the use of BNCT with BPA should be explored in a stratified randomized phase II trial in which patients with the unmethylated MGMT DNA-repair gene are offered BNCT vs. RT plus TMZ.


Ancestry inference using principal component analysis and spatial analysis: a distance-based analysis to account for population substructure.
Accurate inference of genetic ancestry is of fundamental interest to many biomedical, forensic, and anthropological research areas. Genetic ancestry memberships may relate to genetic disease risks. In a genome association study, failing to account for differences in genetic ancestry between cases and controls may also lead to false-positive results. Although a number of strategies for inferring and taking into account the confounding effects of genetic ancestry are available, applying them to large studies (tens thousands samples) is challenging. The goal of this study is to develop an approach for inferring genetic ancestry of samples with unknown ancestry among closely related populations and to provide accurate estimates of ancestry for application to large-scale studies. In this study we developed a novel distance-based approach, Ancestry Inference using Principal component analysis and Spatial analysis (AIPS) that incorporates an Inverse Distance Weighted (IDW) interpolation method from spatial analysis to assign individuals to population memberships. We demonstrate the benefits of AIPS in analyzing population substructure, specifically related to the four most commonly used tools EIGENSTRAT, STRUCTURE, fastSTRUCTURE, and ADMIXTURE using genotype data from various intra-European panels and European-Americans. While the aforementioned commonly used tools performed poorly in inferring ancestry from a large number of subpopulations, AIPS accurately distinguished variations between and within subpopulations. Our results show that AIPS can be applied to large-scale data sets to discriminate the modest variability among intra-continental populations as well as for characterizing inter-continental variation. The method we developed will protect against spurious associations when mapping the genetic basis of a disease. Our approach is more accurate and computationally efficient method for inferring genetic ancestry in the large-scale genetic studies.


Utilization of an interorganizational network analysis to evaluate the development of community capacity among a community-academic partnership.
Following a community health assessment the Brazos Valley Health Partnership (BVHP) organized to address fragmentation of services and local health needs. This regional partnership employs the fundamental principles of community-based participatory research, fostering an equitable partnership with the aim of building community capacity to address local health issues. This article describes changes in relationships as a result of capacity building efforts in a community-academic partnership. Growth in network structure among organizations is hypothesized to be indicative of less fragmentation of services for residents and increased capacity of the BVHP to collectively address local health issues. Each of the participant organizations responded to a series of questions regarding its relationships with other organizations. Each organization was asked about information sharing, joint planning, resource sharing, and formal agreements with other organizations. The network survey has been administered 3 times between 2004 and 2009. Network density increased for sharing information and jointly planning events. Growth in the complexity of relationships was reported for sharing tangible resources and formal agreements. The average number of ties between organizations as well as the strength of relationships increased. This study provides evidence that the community capacity building efforts within these communities have contributed to beneficial changes in interorganizational relationships. Results from this analysis are useful for understanding how a community partnership's efforts to address access to care can strengthen a community's capacity for future action. Increased collaboration also leads to new assets, resources, and the transfer of knowledge and skills.


Biofabrication with biopolymers and enzymes: potential for constructing scaffolds from soft matter.
Regenerative medicine will benefit from technologies capable of fabricating soft matter to have appropriate architectures and that provide the necessary physical, chemical and biological cues to recruit cells and guide their development. The goal of this report is to review an emerging set of biofabrication techniques and suggest how these techniques could be applied for the fabrication of scaffolds for tissue engineering. Electrical potentials are applied to submerged electrodes to perform cathodic and anodic reactions that direct stimuli-responsive film-forming polysaccharides to assemble into hydrogel films. Standard methods are used to microfabricate electrode surfaces to allow the electrical signals to be applied with spatial and temporal control. The enzymes mushroom tyrosinase and microbial transglutaminase are used to catalyze macromolecular grafting and crosslinking of proteins. Electrodeposition of the polysaccharides chitosan and alginate allow hydrogel films to be formed in response to localized electrical signals. Co-deposition of various components (e.g., proteins, vesicles and cells), and subsequent electrochemical processing allow the physical, chemical and biological activities of these films to be tailored. Enzymatic processing allows for the generation of stimuli-responsive protein conjugates that can also be directed to assemble in response to imposed electrical signals. Further, enzyme-catalyzed crosslinking of gelatin allows replica molding of soft matter to create hydrogel films with topological structure. Biofabrication with biological materials and mechanisms provides new approaches for soft matter construction. These methods may enable the formation of tissue engineering scaffolds with appropriate architectures, assembled cells, and spatially organized physical, chemical and biological cues.


[Present status of the health care system].
Characteristic feature of the aggression to BiH are attacks to medical facilities, equipment and staff, as well as to the patients in hospitals. All international conventions were broken, as well as all elementary human rights. Organisation of the work in all medical institutions was spontaneous. Nevertheless, continuity and availability of health care delivery was satisfactory. Many governmental and non-governmental organizations have helped health care system during the war. During the war number of health care institutions increased for several reasons: a) founding 86 points of humanitarian organizations; b) founding 19 temporary hospitals; c) founding new 14 homes of health. Due to war destruction, it is necessary to determine the priorities in sanation of certain segments of health care system.


CathaCyc, a metabolic pathway database built from Catharanthus roseus RNA-Seq data.
The medicinal plant Madagascar periwinkle (Catharanthus roseus) synthesizes numerous terpenoid indole alkaloids (TIAs), such as the anticancer drugs vinblastine and vincristine. The TIA pathway operates in a complex metabolic network that steers plant growth and survival. Pathway databases and metabolic networks reconstructed from 'omics' sequence data can help to discover missing enzymes, study metabolic pathway evolution and, ultimately, engineer metabolic pathways. To date, such databases have mainly been built for model plant species with sequenced genomes. Although genome sequence data are not available for most medicinal plant species, next-generation sequencing is now extensively employed to create comprehensive medicinal plant transcriptome sequence resources. Here we report on the construction of CathaCyc, a detailed metabolic pathway database, from C. roseus RNA-Seq data sets. CathaCyc (version 1.0) contains 390 pathways with 1,347 assigned enzymes and spans primary and secondary metabolism. Curation of the pathways linked with the synthesis of TIAs and triterpenoids, their primary metabolic precursors, and their elicitors, the jasmonate hormones, demonstrated that RNA-Seq resources are suitable for the construction of pathway databases. CathaCyc is accessible online (http://www.cathacyc.org) and offers a range of tools for the visualization and analysis of metabolic networks and 'omics' data. Overlay with expression data from publicly available RNA-Seq resources demonstrated that two well-characterized C. roseus terpenoid pathways, those of TIAs and triterpenoids, are subject to distinct regulation by both developmental and environmental cues. We anticipate that databases such as CathaCyc will become key to the study and exploitation of the metabolism of medicinal plants.


Contrast-Enhanced Endoscopic Ultrasonography for Pancreatic Tumors.
To investigate the usefulness of contrast-enhanced endoscopic ultrasonography (CE-EUS) for histological differentiation of pancreatic tumors. CE-EUS was performed for consecutive patients having a pancreatic solid lesion, and tumors were classified into three vascular patterns (hypervascular, isovascular, and hypovascular) at two time phases (early-phase and late-phase). Correlation between vascular patterns and histopathology of resected pancreatic cancer (PC) tissues was ascertained. The final diagnoses of 147 examined tumors were PC (n = 109), inflammatory mass (n = 11), autoimmune pancreatitis (n = 9), neuroendocrine tumor (n = 8), and others (n = 10). In late-phase images, 104 of 109 PCs had the hypovascular pattern, for a diagnostic sensitivity and specificity of 94% and 71%, respectively. Of 28 resected PCs, 10 had isovascular, and 18 hypovascular, patterns on the early-phase image. Early-phase isovascular PCs were more likely to be differentiated than were early-phase hypovascular PCs (6 well and 4 moderately differentiated versus 3 well, 14 moderately, and 1 poorly differentiated, P = 0.028). Immunostaining revealed that hypovascular areas of early-phase images reflected heterogeneous tumor cells with fibrous tissue, necrosis, and few vessels. CE-EUS could be useful for distinguishing PC from other solid pancreatic lesions and for histological differentiation of PCs.


Trait Associations in Diversity Panels of the Two Common Bean (Phaseolus vulgaris L.) Gene Pools Grown under Well-watered and Water-Stress Conditions.
Common beans are a warm-season, food legume cultivated in areas prone to water limitation throughout their growing season. This study assessed the magnitude and pattern of trait associations for a total of 202 common bean genotypes divided into panels of 81 Andean and 121 Mesoamerican gene pool accessions grown under contrasting treatments of well-watered, non-stress, and water-limited, terminal drought-stress conditions. Linear correlation, complex path coefficient, and genetic divergence analyses were used to dissect the relationship dynamics between traits and the relative contribution of adaptive traits to differentiation among gene pools and genotypes based on drought stress. Drought severity level for the trial was high and created the ideal condition to reveal genotypic differences, as seen by the differential response of the genotypes for the various traits measured. The value for phenotypic coefficients of variation for all traits was higher than the corresponding genotypic values. Seed yield had positive and strong genotypic and phenotypic correlation with pods per plant across gene pools and stress levels. The overall amount of genetic correlation was greater than the corresponding phenotypic correlation matrix for all the traits within the gene pool and across stress levels. Moreover, the results depicted the phenotypic correlation as equal or better than its genotypic counterpart in estimating drought tolerance in common bean plants. Clustering analysis with Mahanalobis's coefficient of generalized distance grouped genotypes with a differential level of drought adaptation into different classes within each panel. This indicates drought tolerance involves different mechanisms of plant response and is present separately in each gene pool panel. Pods per plant, seed weight, pod partitioning index, and harvest index are useful selection objectives to improve drought adaptation in common bean, but must be differentially weighted in each gene pool. The analysis of genetic variation and association between adaptive traits on the two panels provided useful insights on which traits could be used to improve common bean adaptation to low water availability during the growth season.


[Clinical usefulness of alpha-and beta-defensins in biological environments in patients with ulcerative colitis].
To determine the dependence of alpha-and beta-defensins in biological environments and prevalence of inflammatory process, activity bei patienten ulcerative colitis (UC). MATERIALIEN UND METHODEN: The study was conducted on 63 patients with moderate activity UC and 25 practically healthy people. Depending on the extent of the gut was isolated by the three groups of patients: group 1 (n = 18)--distal form, group 2 (n = 24)--left-hand localization of pathological process and group 3 (n = 21)--total form UC. Content of beta-defensins in the stool and alpha-defensins in blood by method of the immune-enzyme analysis. In the control group the level of beta-defensins in the stool was 3.7 +/- 3.2 ng/g, alpha-defensins in blood--71.7 +/- 3.8 ng/ml. In patients of group 1 the concentration of beta-defensins in the stool was 218.5 +/- 8.2 ng/g in 2 group--210.8 +/- 4.3 ng/g and in group 3--261.7 +/- 9.2 ng/g. Intermediate level a-defensins in blood match in group 1--471.8 ğ 9.9 ng/ml in group 2--853.5 ğ 9.5 ng/ml and in group 3--479.3 +/- 6.8 ng/ ml. Regardless of the localization of lesions in the gut, not including beta-defensin in the stool at 2 under, the concentration of antimicrobial peptides in remission. Proven association increased expression of alpha-and beta-defensins with activity and a length of inflammation in the intestine at UC.


Coupling Machine Learning Models with Innovative Technology-based Screening Tool for Identifying Psychological Distress among Aboriginal Perinatal Mothers.
Aboriginal perinatal mothers are at a significant risk of experiencing mental health problems, which can have profound negative impacts, despite their overall resilience. This work aimed to build prediction models for identifying high psychological distress among Aboriginal perinatal mothers by coupling machine learning models with an innovative and culturally-safe screening tool. The original dataset of 179 Aboriginal mothers with 337 variables was obtained from twelve perinatal health settings at Perth metropolitan and regional centers in Western Australia between July and September 2022, using a specifically designed web-based rubric for the perinatal mental health assessment. After data preprocessing and feature selection, 23 variables related to emotional manifestations, the problematic partner, worries about daily living, and the need for follow-up wraparound support were identified as significant predictors for the high risk of psychological distress measured by the Kessler 5 plus adaptation. The selected predictors were used to train prediction models, and most of the chosen machine learning models achieved satisfactory results, with Random Forest and Support Vector Machine yielding the highest AUC of over 0.95, accuracy over 0.86, and F1 score above 0.87. This study demonstrates the potential of using machine learning-based models in clinical decision-making to facilitate healthcare and social and emotional well-being for Aboriginal families.


Jasmonic acid is induced in a biphasic manner in response of pea seedlings to wounding.
The role of jasmonic acid (JA) in plant wounding response has been demonstrated. However, the source of JA in wound signaling remains unclear. In the present study, pea seedlings were used as material to investigate the systemic induction of JA and the activation of lipoxygenase (LOX)-dependent octadecanoid pathway upon wounding. The results showed that endogenous JA could induce two peaks in the wounded leaves and the stalks, while only one peak in the systemic leaves. LOX activity and its protein amount were also induced and the stimulation mainly occurred in the late phase, while one peak of induction was present after pretreatment with JA. Applied nordihydroguaiaretic acid (NDGA), an inhibitor of LOX activity, only inhibited the induction of JA in the late phase, and the resistance of pea was impaired. Furthermore, 13(S)-hydroperoxy-9(Z), 11(E)-octadecadienoic acid (13(S)-H(P)ODE) was confirmed to be the main product of LOX throughout the experimental time. In addition, immunocytochemical analysis also revealed the occurrence of JA biosynthesis and transport upon wounding. These results demonstrated that wound-induced JA in wounded leaves resulted from its biosynthesis and conversion from its conjugates, while in systemic leaves resulted from its transport and biosynthesis; and proved that the LOX pathway was vital to the wound-induced defense response involved in JA biosynthesis.


Assessment of epiretinal membrane formation using en face optical coherence tomography after rhegmatogenous retinal detachment repair.
To investigate epiretinal membrane (ERM) formation using en face optical coherence tomography (OCT) after vitrectomy for rhegmatogenous retinal detachment (RRD). We retrospectively reviewed the medical records of 64 consecutive eyes (64 patients) with RRD treated by vitrectomy without ERM and internal limiting membrane peeling. ERMs and retinal folds were detected by B-scan and en face imaging. The maximum depth of retinal folds (MDRF) was quantified using en face imaging. ERM severity was staged using B-scan imaging. Main outcome measures were ERM detection rate with B-scan and en face imaging, MDRF, ERM staging, postoperative best-corrected visual acuity (BCVA; logarithm of the minimum angle of resolution), and risk factors for ERM formation. The detection rate for ERM formation was significantly higher with en face imaging (70.3%) than with B-scan imaging (46.9%; P = 0.007). There was no significant difference in postoperative BCVA between eyes with ERM formation (0.06 ± 0.26) and those without ERM formation (0.01 ± 0.14; P = 0.298). Forty of 45 (88.9%) eyes with ERM formation were classified as stage 1. Twenty-seven of 45 (60.0%) eyes with ERM formation developed parafoveal retinal folds. The mean MDRF was 27.4 ± 32.2 μm. Multiple retinal breaks and a maximum retinal break size of ≥ 2 disc diameters were significantly associated with ERM formation (P = 0.033 and P = 0.031, respectively). Although ERM formation was observed in 70.3% patients after RRD repair, the formed ERM was not severe and had minimal impact on the postoperative visual acuity.


Glycine decreases desensitization of N-methyl-D-aspartate (NMDA) receptors expressed in Xenopus oocytes and is required for NMDA responses.
In Xenopus oocytes injected with rat brain mRNA, as in neurons, glycine greatly potentiated responses of the N-methyl-D-aspartate (NMDA) type of excitatory amino acid receptor. Injected oocytes generated a partially desensitizing inward current in response to NMDA with 30 nM added glycine. As the added glycine concentration was increased from 30 nM to 1 microM, the NMDA response was increased and exhibited less desensitization. The relationship between the NMDA peak response and added glycine concentration indicated a single component response with apparent affinity of 0.29 microM and a Hill coefficient of 0.77. The desensitized response was also fit by the Hill relation with a lower affinity but similar coefficient. The time course of desensitization at 500 microM NMDA was exponential with a time constant (350 msec) that was independent of glycine concentration between 0.03 and 0.3 microM. At higher glycine concentration a slower component of decay (tau = 1.4 sec) was observed. This component was enhanced by increasing the extracellular Ca2+. NMDA without added glycine evoked a small transient response. However this response was suppressed completely by prewashing with the glycine antagonist 7-chlorokynurenic acid, suggesting that it may have been due to glycine contamination. The dose-response relation for low concentrations of glycine indicated that the measured level of glycine contamination accounted for these responses. These results indicate that glycine has at least two actions at the NMDA receptor: it enables channel opening by the agonist and decreases desensitization.


32-Channel transmit beamformer with high timing resolution for high-frequency ultrasound imaging systems.
This paper presents a 32-channel high timing resolution transmit-beamforming circuit for use in high-frequency ultrasound imaging systems. Conventional transmit-beamforming circuits are typically implemented using field-programmable gate array (FPGA) chips. However, it is difficult for FPGAs to provide high timing resolution to meet the beamforming requirements of high-frequency ultrasound imaging systems. The proposed transmit-beamforming design can generate stable and suitable delays to excite 32-channel array transducer elements without variations in the process, voltage, and temperature. In addition, the proposed low-complexity architecture can maintain the duty cycle of long prorogation signals with low hardware cost to meet the timing requirements of a large channel number array transducer. The proposed designed transmit beamformer uses 0.18-µm CMOS technology for a 30-MHz high-frequency linear array, and the simulation results show that the proposed transmit-beamforming application-specific integrated circuit can achieve a maximum time delay of 619.5 ns with a time resolution of 617 ps.


Cytokinin-induced mRNAs in cultured soybean cells.
We have isolated 20 cDNA clones for which the corresponding mRNAs accumulate 2- to 20-fold within 4 hr of cytokinin addition to cytokinin-starved soybean suspension cultures. These changes in mRNA levels precede the greening and cell growth induced by cytokinin in this system. Treatment with cycloheximide before cytokinin addition enhances the accumulation of these messages. In addition, the abundance of these mRNAs is influenced by auxin. We demonstrate that the mRNAs corresponding to the cDNAs in this collection display a variety of patterns of accumulation in the tissues of an intact plant. We also show that different cytokinins induce similar increases in message levels. However, analogs of cytokinin that are much less active in bioassays induce lower levels of message accumulation. Sequence analyses indicate that two of the cDNAs correspond to ribosomal protein genes. The remaining cDNAs are likely to represent previously undiscovered genes that function early in the initiation of plant cell growth and/or chloroplast development.


A rare pattern of angiogenesis in meningiomas: glomeruloid microvascular proliferation.
Glomeruloid microvascular proliferation (GMP) is a localized proliferation of vascular endothelial cells resembling a renal glomerule. The nature of cells participating in the formation of these structures remains unclear. While it is a characteristic feature of glioblastoma, it is rarely seen in other solid tumors. Presence of diffuse GMP in meningiomas is characterized by peritumoral edema and an atypical contrast uptake in radiological imaging. Due to its rare nature, a case of spinal meningioma comprising distinct GMP was presented in this study in company with literature data. Also provided a discussion on the pathogenesis of this unusual pattern of angiogenesis and its relationship with tumors biological behavior.


Carbapenemase-producing Enterobacteriaceae and non-Enterobacteriaceae from animals and the environment: an emerging public health risk of our own making?
Acquired carbapenemases pose one of the most pressing public health threats relating to antibiotic resistance. In most countries, the number of carbapenemase-producing bacteria from human clinical specimens is rising, and the epidemiological status of these multiresistant bacteria is progressively worsening. Furthermore, there is a growing number of reports of carbapenemases found either in bacteria isolated from non-human sources or in Salmonella enterica subsp. enterica, a zoonotic species. However, carbapenemases are not yet systematically sought in bacteria from non-human sources, reports of them are largely observational, and there is limited investigation of carbapenemase-positive bacteria in animals and possible links with people who may have acted as potential sources. Active surveillance and monitoring for carbapenem-resistant bacteria in the food chain and other non-human sources is urgently needed, with an enhanced and rigorous follow-up of all positive results. The carbapenems are currently our last good defence against multiresistant Gram-negative bacteria. Our ability to limit the rise and spread of carbapenemase producers, which occur only at basal levels in many countries at present, should serve as a key performance indicator for the success or failure of the efforts that have been called for by international organizations and governments to reduce the impact of antibiotic resistance.


[Quantitative assessment of antithymocyte antibodies in schizophrenia].
The use of the method of fluorescent antibodies and cytotoxic tests showed that the blood serum of schizophrenic patients and normals possess an antithymocytic activity. However, the level of this activity in patients exceeds the corresponding indices in normals (p less than 0.02). The use of indirect immunofluorescent methods permitted to demonstrate that the antithymic antibody titres in schizophrenic patients is much higher than in normals. The amount of the cytotoxic index is in direct correlation with the level of antithymic antibodies determined by the fluorescent antibody method.


Courtship dives of Anna's hummingbird offer insights into flight performance limits.
Behavioural displays are a common feature of animal courtship. Just as female preferences can generate exaggerated male ornaments, female preferences for dynamic behaviours may cause males to perform courtship displays near intrinsic performance limits. I provide an example of an extreme display, the courtship dive of Anna's hummingbird (Calypte anna). Diving male Anna's hummingbirds were filmed with a combination of high-speed and conventional video cameras. After powering the initial stage of the dive by flapping, males folded their wings by their sides, at which point they reached an average maximum velocity of 385 body lengths s(-1) (27.3 m s(-1)). This is the highest known length-specific velocity attained by any vertebrate. This velocity suggests their body drag coefficient is less than 0.3. They then spread their wings to pull up, and experienced centripetal accelerations nearly nine times greater than gravitational acceleration. This acceleration is the highest reported for any vertebrate undergoing a voluntary aerial manoeuvre, except jet fighter pilots. Stereotyped courtship behaviours offer several advantages for the study of extreme locomotor performance, and can be assessed in a natural context.


Use of Health Behavior Theory in Funded Grant Proposals: Cancer Screening Interventions as a Case Study.
Interventions using theory should change behavior and identify both mechanisms of effect and necessary conditions. To date, inconsistent description of "use of theory" has limited understanding of how theory improves intervention impact. The purpose of this study was to describe the use of theory in health behavior intervention development by coding grant proposals. We developed an abstraction tool to characterize investigators, interventions, and theory use and identified seven core elements describing both how and how much theory was used. We used the tool to review and code NCI's funded cancer screening intervention R01 proposals, 1998-2009. Of 116 proposals, 38 met criteria; all but one described a conceptual model unique to the proposed research. Few proposals included plans to identify mechanisms of effect or conditions necessary for intervention effectiveness. Cancer screening intervention grant proposals rarely use theory in ways that advance behavioral or theoretical sciences. Proposed core elements may classify and synthesize the use of theory in behavioral intervention research.


Inducible anti-sense RNA for angiotensinogen in stably transformed hepatoma cell lines.
Angiotensinogen mRNA is found in many extrahepatic tissues, where it may participate in local angiotensin-generating systems. In this study we explore the feasibility of using anti-sense RNA to decrease angiotensinogen production in rat H4IIEC3 hepatoma cells. An amplifiable shuttle vector was modified to allow the production of high levels of stable anti-sense RNA from two regions of the mouse angiotensinogen gene under the control of the inducible sheep metallothionein promoter. Stably transformed, clonal cell lines expressing anti-sense RNA for angiotensinogen were isolated after selection with the aminoglycoside G418. Subsequently, the number of chromosomally integrated copies of the angiotensinogen anti-sense constructs was coamplified by methotrexate selection for dihydrofolate reductase activity carried on the shuttle vector. With a 20- to 30-fold induction of the anti-sense RNAs, the target angiotensinogen mRNA level was reduced to 25-30% of control values. The specificity of this effect was confirmed by showing no decrease in either beta-tubulin or neomycin phosphotransferase mRNA levels. Using tissue-specific promoters, it should be possible to direct these effects to specific organs in transgenic mice. However, in agreement with results from other groups, our findings suggest that it will not be possible to eradicate completely the target gene product using the anti-sense RNA strategy.


Surgical management of ovarian cancer.
Ovarian cancer represents the second most common cancer of the female reproductive organs. It is usually not associated with significant warning signals that led to early diagnosis. The management is based on adequate surgical staging, removal of as much of the bulk tumor as possible, careful histologic assessment of the primary cancer and its metastases, and appropriate postoperative adjuvant therapy. Stage I cancer confined to one ovary and with a very favorable histology, such as borderline malignant epithelial cancers, dysgerminoma, granulosa cell tumor, and Sertoli-Leydig's cell tumor, may be treated with surgery alone. Stage I epithelial cancers are best treated with a total abdominal hysterectomy, bilateral salpingo-oophorectomy, and adjuvant chemotherapy or radiation therapy. Stage I nonepithelial malignant neoplasms are best treated with a unilateral salpingo-oophorectomy followed by an aggressive combination chemotherapy. Advanced ovarian cancers regardless of the histologic origin of the tissue should be treated with aggressive surgery, including complete removal of the tumor whenever possible followed by adjuvant therapy selected on the basis of the histologic appearance of the tumor. Complete remissions for ovarian cancer should be confirmed by second-look operation prior to discontinuing adjuvant chemotherapy.


Glycoalkaloids of wild and cultivated Solanum: effects on specialist and generalist insect herbivores.
Plant domestication by selective breeding may reduce plant chemical defense in favor of growth. However, few studies have simultaneously studied the defensive chemistry of cultivated plants and their wild congeners in connection to herbivore susceptibility. We compared the constitutive glycoalkaloids (GAs) of cultivated potato, Solanum tuberosum, and a wild congener, S. commersonii, by liquid chromatography coupled to mass spectrometry. We also determined the major herbivores present on the two species in field plots, and tested their preference for the plants and their isolated GAs in two-choice bioassays. Solanum commersonii had a different GA profile and higher concentrations than S. tuberosum. In the field, S. tuberosum was mostly attacked by the generalist aphids Myzus persicae and Macrosiphum euphorbiae, and by the specialist flea beetle Epitrix argentinensis. In contrast, the most common herbivore on S. commersonii was the specialist sawfly Tequus sp. Defoliation levels were higher on the wild species, probably due to the chewing feeding behavior of Tequus sp. As seen in the field, M. persicae and E. argentinensis preferred leaf disks of the cultivated plant, while Tequus sp. preferred those of the wild one. Congruently, GAs from S. commersonii were avoided by M. persicae and preferred by Tequus sp. The potato aphid performed well on both species and was not deterred by S. commersonii GAs. These observations suggest that different GA profiles explain the feeding preferences of the different herbivores, and that domestication has altered the defensive capacity of S. tuberosum. However, the wild relative is still subject to severe defoliation by a specialist herbivore that may cue on the GAs.


Understanding the link between pneumococcal serotypes and invasive disease.
To determine the relationship between serotypes and the burden of pneumococcal disease in Latin American children, population- and laboratory-based surveillance data for the region were evaluated. Streptococcus pneumoniae continues to be an important bacterial pathogen causing infections in children. A seven-valent pneumococcal conjugate vaccine (PCV7) has been documented to be extremely effective in the prevention of invasive and non-invasive S. pneumoniae infections due to vaccine serotypes. However, due to evolving epidemiology, there is a need to enhance S. pneumoniae serotype coverage in future generation vaccines, complemented by ongoing surveillance of pneumococcal serotypes following vaccine introduction.


[Clinical characteristics of post-stroke anxiety disorders].
Authors studied 198 patients with first-ever cerebral stroke. Affective disorders, including depression, generalized anxiety disorder (GAD) and phobias, were identified in 71.2% of patients. The study demonstrated that the anxiety syndrome developed in the post-stroke period may be regarded as relatively independent. Some post-stroke anxiety disorders had different risk and pathogenetic factors. GAD is associated with young age and phobias are associated with female sex and inherited predisposition. In contrast to post stroke depression, anxiety-phobic disorders develop more frequently in patients with mild stroke.


Surgery for acquired valvular heart disease: the San Pablo Heart Institute experience.
Ninety-nine patients underwent surgery for acquired diseases of the mitral and aortic valves between August 22, 1988 and August 21, 1990, of these, 64 procedures were done on the aortic valve and 35 procedures on the mitral valve. The early mortality for mitral valve surgery was 5.8% and late death occurred at a rate of 8.8%. As for the aortic valve surgery the operative mortality was zero and late death occurred at a rate of 3.4%. Thromboembolism occurred at a rate of 3.3% per patient year for patient undergoing mitral valve surgery and 6.3% per patient year for those undergoing aortic valve surgery. Valve thrombosis occurred at a rate of 6.6% per patient year for patients undergoing mitral valve surgery and zero for patients undergoing aortic valve surgery. Anticoagulation related bleeding occurred at a rate of 49% per patient year for patients undergoing mitral valve surgery and fatal bleeding at a rate of 3.3% per patient year. For those patients undergoing aortic valve surgery bleeding occurred at a rate of 25% per patient year and fatal bleeding occurred at a rate of 3.1%. This rate of bleeding appears high; and is due to poorly monitored anticoagulation.


Annual summary of vital statistics--1996.
Several recent trends in the vital statistics of the United States continued in 1996, including an increase in life expectancy and declines in infant mortality, births to teenage mothers, age-adjusted death rates, and death rates for children and adolescents. In 1996, there were an estimated 3 914 953 births in the United States. The preliminary birth rate remained unchanged at 14.8 births per 1000 population, and the fertility rate, births per 1000 women 15 to 44 years of age, was essentially the same at 65.7. Fertility rates rose slightly for most racial and ethnic groups except black women, for whom the rate hit a historic low of 70.8. Overall, fertility remains particularly high for Hispanic women, although there is considerable variation within this heterogenous group. For the fifth consecutive year, birth rates dropped for teenagers. Birth rates for women >/=30 years of age continued to increase. The birth rate for unmarried women declined 1% in 1996 to 44.6 births per 1000 unmarried women, continuing the decline noted in 1995 for the first time in 2 decades. The percentage of women who began prenatal care in the first trimester rose in 1996 to 81.8%, whereas the percentage with late (third trimester) or no care dropped to 4.1%. The rise in timely prenatal care was greatest for black and Hispanic women. The percentage of low birth weight (LBW) infants reached 7.4% in 1996, its highest level since 1975. The very low birth weight rate remained unchanged at 1.4%. The rise in LBW occurred primarily among white women, whereas the LBW rate for black women dropped to 13.0%, the lowest rate reported since 1987. The rise among white women is only partially a result of increases in multiple births, because LBW rates have also risen among white singleton births. The multiple birth ratio rose again in 1996 by 2%, as it has since 1980. The rise was particularly large for higher-order multiple births. Infant mortality reached an all time low level of 7.2 deaths per 1000 births, based on preliminary 1996 data. Neonatal and postneonatal rates declined, as did rates for both black and white infants. National birth weight specific mortality rates are reported here for the first time. In 1995, 63% of infant deaths occurred to the 7.3% of the population that was born LBW. The four leading cause of infant death were congenital anomalies, disorders relating to short gestation and unspecified birth weight, sudden infant death syndrome, and respiratory distress syndrome, accounting for more than half of infant deaths in 1996. Despite the declines in infant mortality, the United States continues to rank poorly in international comparisons of infant mortality. Expectation of life at birth reached a new high in 1996 of 76.1 years for all gender and race groups combined. Age-adjusted mortality rates declined in 1996 for diseases of the heart, malignant neoplasms, cerebrovascular diseases, accidents and adverse effects, chronic liver disease and cirrhosis, and suicide. They rose, as in the past several years, for chronic obstructive pulmonary diseases, diabetes mellitus, and pneumonia and influenza. For the first time since human immunodeficiency virus infection was created as a special cause-of-death category in 1987, death rates for human immunodeficiency virus infection declined from 15.6 in 1995 to 11.6 in 1996. The homicide rate also declined, as it has since 1991. Death rates for children between 1 and 19 years of age declined in 1996, with an estimated 29 183 deaths to children. Unintentional injury mortality has dropped by approximately 50% among children and adolescents since 1979, although it remains the leading cause of death for all age groups of children from 1 to 19 years. Homicide was the fourth leading cause of death for children 1 to 4 and 5 to 9 years of age, the third leading cause for children 10 to 14, and the second leading cause for 15 to 19 year olds.


CD200R1 and CD200R1L expression is regulated during B cell development in swine and modulates the Ig production in response to the TLR7 ligand imiquimoid.
The CD200R family comprises a group of paired receptors that can modulate the activation of immune cells. They are expressed both on myeloid cells and lymphocyte subsets. Here we report that the expression of these receptors on porcine B cells is tightly regulated, being mainly expressed on mature cells. The expression of the inhibitory receptors CD200R1 and/or its splicing variant CD200R1X2, either in combination or not with the activating receptor CD200R1L, is upregulated in sIgM+ effector/memory cells, and tends to decline thereafter as these cells progress to plasmablasts or switch the Ig isotype. sIgM+ naïve and primed cells only express, by contrast, the CD200R1X2 receptor. B-1 like cells also express CD200R1 isoforms, either alone or in combination with CD200R1L. Treatment of peripheral blood mononuclear cells with a monoclonal antibody specific for inhibitory receptors, enhances the IgM and IgG production induced by TLR7 stimulation suggesting a modulatory role of B cell functions of these receptors.


Variation in market access decisions for cell and gene therapies across the United States, Canada, and Europe.
Transformative cell and gene therapies have now launched worldwide, and many potentially curative cell and gene therapies are in development, offering the prospect of significant health gains for patients. Access to these therapies depend on decisions made by health technology assessment (HTA) and payer organizations. We sought to describe the emerging cell and gene therapies market access landscape by analyzing 17 US commercial payer medical policies, and HTA reports from five European countries and Canada. We found that some US health plans applied coverage restrictions more often than others (four plans applied restrictions in all decisions, while four plans applied restrictions in <30% of decisions). The European and Canadian HTA bodies recommend access to fewer therapies than US health plans, reflecting a more stringent approach in the context of limited evidence and high scientific uncertainty that is commonly associated with these treatments. Our findings suggest that patient access to approved cell and gene therapies is restricted in all regions studied, though the nature of these restrictions differs between US health plans and the European/Canada HTA recommendations. Payers, HTA groups, pharmaceutical companies, and other stakeholders should collaborate to more clearly define the "uncertainties" and develop market access policies that balance benefits of early access with ongoing data collection to close evidence gaps over time.


Temporal control of gene deletion in sensory ganglia using a tamoxifen-inducible Advillin-Cre-ERT2 recombinase mouse.
Tissue-specific gene deletion has proved informative in the analysis of pain pathways. Advillin has been shown to be a pan-neuronal marker of spinal and cranial sensory ganglia. We generated BAC transgenic mice using the Advillin promoter to drive a tamoxifen-inducible CreERT2 recombinase construct in order to be able to delete genes in adult animals. We used a floxed stop ROSA26LacZ reporter mouse to examine functional Cre expression, and analysed the behaviour of mice expressing Cre recombinase. We used recombineering to introduce a CreERT2 cassette in place of exon 2 of the Advillin gene into a BAC clone (RPCI23-424F19) containing the 5' region of the Advillin gene. Transgenic mice were generated using pronuclear injection. The resulting AvCreERT2 transgenic mice showed a highly specific expression pattern of Cre activity after tamoxifen induction. Recombinase activity was confined to sensory neurons and no expression was found in other organs. Less than 1% of neurons showed Cre expression in the absence of tamoxifen treatment. Five-day intraperitoneal treatment with tamoxifen (2 mg per day) induced Cre recombination events in ≈90% of neurons in dorsal root and cranial ganglia. Cell counts of dorsal root ganglia (DRG) from transgenic animals with or without tamoxifen treatment showed no neuronal cell loss. Sensory neurons in culture showed ≈70% induction after 3 days treatment with tamoxifen. Behavioural tests showed no differences between wildtype, AvCreERT2 and tamoxifen-treated animals in terms of motor function, responses to light touch and noxious pressure, thermal thresholds as well as responses to inflammatory agents. Our results suggest that the inducible pan-DRG AvCreERT2 deleter mouse strain is a useful tool for studying the role of individual genes in adult sensory neuron function. The pain phenotype of the Cre-induced animal is normal; therefore any alterations in pain processing can be unambiguously attributed to loss of the targeted gene.


An alternative pathway for ureide usage in legumes: enzymatic formation of a ureidoglycolate adduct in Cicer arietinum and Phaseolus vulgaris.
Ureidoglycolate is an intermediate in the degradation of the ureides, allantoin and allantoate, found in many organisms. In some leguminous plant species these compounds are used to transport recently fixed nitrogen in the root nodules to the aerial parts of the plant. In the present study, it was demonstrated that purified ureidoglycolases from chickpea (Cicer arietinum) and French bean (Phaseolus vulgaris) do not produce glyoxylate, and can use phenylhydrazine as a substrate with K(m) values of 4.0 mM and 8.5 mM, respectively. Furthermore, these enzymes catalyse the transfer of the ureidoglycolyl group to phenylhydrazine to produce ureidoglycolyl phenylhydrazide, which degrades non-enzymatically to glyoxylate phenylhydrazone and urea. This supports their former classification as ureidoglycolate urea-lyases. The enzymatic reaction catalysed by the characterized ureidoglycolases uncovered here can be viewed as a novel type of phenylhydrazine ureidoglycolyl transferase. The implications of these findings for ureide metabolism in legume nitrogen metabolism are discussed.


Ameloblastic Fibrosarcoma of the Jaw: Case Report, Genetic Profiling, and Literature Review.
Ameloblastic fibrosarcoma (AFS) is considered a malignant progression resulting from dysplastic changes in an ameloblastic fibroma (AF). Both tumors are extremely rare, with only a few cases reported in the scientific literature. Notably, BRAF mutations have been identified in ameloblastomas, suggesting a connection between ameloblastic morphology and BRAF mutations, as AF is believed to be the precursor neoplasm leading to AFS. In this study, we present a case of AFS in a 25-year-old male. The tumor tissue underwent molecular analysis, specifically next-generation sequencing (NGS) using the Oncomine Comprehensive Assay v3 System. The analysis revealed pathogenic mutations in TP53 and RB genes, as well as copy number gains in NTRK1, MDM4, and BRAF. Additionally, we provide a summary of the literature's findings from the analysis of 107 previously reported AFS cases. Our findings suggest the existence of a molecularly distinct subtype, emphasizing the importance of comprehensive molecular testing for these patients.


[Image-guided percutaneous gastrostomy].
Gastrostomy is mainly used to provide longterm enteral nutrition. Percutaneous techniques are generally preferred to surgery except for specific cases. Image-guided percutaneous gastrostomy, currently used less than the gastroscopy-guided technique, is a simple, reliable and advantageous technique in managing these frequently debilitated patients. The different aspects of the procedure will be described: indications, contraindications, technique, follow-up, main complications and technical variations.


The family perspective of ESRD deaths.
Nephrologists need to become more aware of how patients die. Families of patients who died after receiving dialysis at 5 New England nephrology clinics were contacted and informed about the study. Postal questionnaires were completed 6 to 10 weeks after deaths to elicit the family members' perspectives. The tool inquired about quality of dying, site of death, advance care preferences, whether dialysis was discontinued, and terminal symptoms. There were 188 deaths, and 86 completed questionnaires (46%) were returned. The majority (64%) of family members believed that the patient had a peaceful death, followed by those who were unsure (24%), and those who thought the death was not peaceful (12%). Most deaths took place in institutional settings (56% in the hospital and 14% in nursing homes), and one quarter (27%) of the deaths occurred at home. Whereas some respondents perceived that patients preferred treatment to primarily extend life (28%), more believed that relieving pain (49%) was of greater importance, and the balance (24%) were unsure. Three quarters of the patient population were observed to be in pain during their last week of life. Pain was assessed as being extremely or moderately severe in more than 90% of patients. The second most distressing symptom was weakness or fatigue, followed by dyspnea, gastrointestinal problems, anxiety, and depression. There was significantly less likelihood of pain in the last week of life for patients who died at home compared with those who died in an institution. Nephrologists need to explore new ways to improve symptomatic treatment and minimize suffering of dying dialysis patients. These findings are the first step in benchmarking the prevalence and severity of terminal symptoms.


Long-term treatment effects of vitamin E for tardive dyskinesia.
Several studies have found that alpha-tocopherol (vitamin E) can effectively treat tardive dyskinesia (TD). A limitation of these trials is their short treatment durations (maximum of 12 weeks), which do not allow us to address the effects of long-term treatment. To participate, patients had to have TD and be on stable oral medications. The study enrolled 40 patients who received up to 36 weeks of treatment with d-vitamin E (1600 IU per day) or placebo. Using the Abnormal Involuntary Movements Scale (AIMS) score (sum of items #1-7) to measure TD severity, the study found a significant difference (3 points) in mean AIMS scores, in favor of vitamin E, starting at 10 weeks of treatment and continuing through the full 36 weeks. We used linear mixed-effects regression to quantify the impact of several covariates, and found that treatment assignment. TD duration, and chlorpromazine equivalents had significant effects on decreasing the AIMS score. The study's finding that vitamin E is effective in treating TD agrees with results from prior studies and provides evidence that the effect may extend to treatment of up to 36 weeks. These findings are in direct contrast to those of VA Cooperative Study #394, a much larger, long-term, multi-site study, conducted by many of the same investigators, in which Vitamin E was not superior to placebo.


The importance of increasing primary vaccinations against COVID-19 in Europe.
In the European Union, mass vaccination against COVID-19 staved off the strict restrictions that had characterized early epidemic response. Now, vaccination campaigns are focusing on booster doses, and primary vaccinations have all but halted. Still, 52 million European adults are unvaccinated. We investigated if reaching the still unvaccinated population in future vaccination campaigns would substantially decrease the current burden of COVID-19, which is substantial. We focused on vaccination homophily, whereby those who are unvaccinated are mostly in contact with other unvaccinated, making COVID-19 circulation easier. We quantified vaccination homophily and estimated its impact on COVID-19 circulation. We used an online survey of 1,055,286 people from 22 European countries during early 2022. We computed vaccination homophily as the association between reported vaccination status and perceived vaccination uptake among one's own social contacts, using a case-referent design and a hierarchical logistic model. We used this information in an analysis of the COVID-19 reproduction ratio to determine the impact of vaccine homophily in transmission. Vaccination homophily was present and strong everywhere: the average odds ratio of being vaccinated for a 10-percentage-point increase in coverage among contacts was 1.66 (95% CI=(1.60, 1.72)). Homophily was positively associated with the strictness of COVID-19-related restrictions in 2020 (Pearson = 0.49, P = .03). In the countries studied, 12%-to-18% of the reproduction ratio would be attributable to vaccine homophily. Reducing vaccination homophily may curb the reproduction ratio substantially even to the point of preventing recurrent epidemic waves. In addition to boosting those already vaccinated, increasing primary vaccination should remain a high priority in future vaccination campaigns, to reduce vaccination homophily: this combined strategy may decrease COVID-19 burden.


Phenylacetyl Coenzyme A, Not Phenylacetic Acid, Attenuates CepIR-Regulated Virulence in Burkholderia cenocepacia.
During phenylalanine catabolism, phenylacetic acid (PAA) is converted to phenylacetyl coenzyme A (PAA-CoA) by a ligase, PaaK, and then PAA-CoA is epoxidized by a multicomponent monooxygenase, PaaABCDE, before further degradation through the tricarboxylic acid (TCA) cycle. In the opportunistic pathogen Burkholderia cenocepacia, loss of paaABCDE attenuates virulence factor expression, which is under the control of the LuxIR-like quorum sensing (QS) system, CepIR. To further investigate the link between CepIR-regulated virulence and PAA catabolism, we created knockout mutants of the first step of the pathway (PAA-CoA synthesis by PaaK) and characterized them in comparison to a paaABCDE mutant using liquid chromatography-tandem mass spectrometry (LC-MS/MS) and virulence assays. We found that while loss of PaaABCDE decreased virulence, deletion of the paaK genes resulted in a more virulent phenotype than that of the wild-type strain. Deletion of either paaK or paaABCDE led to higher levels of released PAA but no differences in levels of internal accumulation compared to the wild-type level. While we found no evidence of direct cepIR downregulation by PAA-CoA or PAA, a low-virulence cepR mutant reverted to a virulent phenotype upon removal of the paaK genes. On the other hand, removal of paaABCDE in the cepR mutant did not impact its attenuated phenotype. Together, our results suggest an indirect role for PAA-CoA in suppressing B. cenocepacia CepIR-activated virulence.IMPORTANCE The opportunistic pathogen Burkholderia cenocepacia uses a chemical signal process called quorum sensing (QS) to produce virulence factors. In B. cenocepacia, QS relies on the presence of the transcriptional regulator CepR which, upon binding QS signal molecules, activates virulence. In this work, we found that even in the absence of CepR, B. cenocepacia can elicit a pathogenic response if phenylacetyl-CoA, an intermediate of the phenylacetic acid degradation pathway, is not produced. Instead, accumulation of phenylacetyl-CoA appears to attenuate pathogenicity. Therefore, we have discovered that it is possible to trigger virulence in the absence of CepR, challenging the classical view of activation of virulence by this QS mechanism. Our work provides new insight into the relationship between metabolism and virulence in opportunistic bacteria. We propose that in the event that QS signaling molecules cannot accumulate to trigger a pathogenic response, a metabolic signal can still activate virulence in B. cenocepacia.


Not-for-profit not equal to no profit: profitability planning in not-for-profit organizations.
Contrary to common understanding, not-for-profit organizations must earn a profit in order to maintain fiscal solvency. This article presents a methodology to determine the appropriate profitability target with illustrations presented for a health maintenance organization.


Vegetarian and Vegan Diets: Beliefs and Attitudes of General Practitioners and Pediatricians in France.
Studies suggest a decreasing trend in the consumption of meat products and a growing interest in vegetarian diets. Medical support may be relevant, especially when switching to a vegan diet. Our objective was to describe the beliefs and attitudes of primary care physicians toward vegetarian diets. A cross-sectional survey was conducted among general practitioners and pediatricians thorough a questionnaire including socio-demographic characteristics, specific care to vegetarians, and the risks and benefits of vegetarian diets according to physicians. Out of the 177 participating physicians, 104 (59%) have seen at least one vegetarian patient in consultation in the last three months. Half of the physicians declared that they would dissuade their patients from switching to a vegan diet (n = 88, 51%) and 14% (n = 24) from switching to an ovo-lacto-vegetarian (OLV) diet. Most physicians (n = 141, 88%) did not feel informed enough about these diets. Physicians thought that the most frequent deficiencies for OLV and vegan diets were iron (76% and 84%, respectively) and protein (45% and 79%, respectively). These results highlight the fact that French primary care physicians feel concerned by this subject and need more information on these diets. Specific recommendations would be useful to support their practice and relationship with vegetarians.


Calcium gradients during excitation-contraction coupling in cat atrial myocytes.
1. Confocal microscopy in combination with the calcium-sensitive fluorescent probe fluo-3 was used to study spatial aspects of intracellular Ca2+ signals during excitation-contraction coupling in isolated atrial myocytes from cat heart. 2. Imaging of [Ca2+]i transients evoked by electrical stimulation revealed that Ca2+ release started at the periphery and subsequently spread towards the centre of the myocyte. 3. Blocking sarcoplasmic reticulum (SR) Ca2+ release with 50 microM ryanodine unmasked spatial inhomogeneities in the [Ca2+]i was higher in the periphery than in central regions of the myocyte. 4. Positive (or negative) staircase or postrest potentiation of the 'whole-cell' [Ca2+] transients were paralleled by characteristic changes in the spatial profile of the [Ca2+]i signal. With low SR Ca2+ load [Ca2+]i transients in the subsarcolemmal space were small and no Ca2+ release in the centre of the cell was observed. Loading of the SR increased subsarcolemmal [Ca2+]i transient amplitude and subsequently triggered further release in more central regions of the cell. 5. Spontaneous Ca2+ release from functional SR units, i.e. Ca2+ sparks, occurred at higher frequency in the subsarcolemmal space than in more central regions of the myocyte. 6. Visualization of the surface membrane using the membrane-selective dye Di-8-ANEPPS demonstrated that transverse tubules (t-tubules) were absent in atrial cells. 7. It is concluded that in atrial myocytes voltage-dependent Ca2+ entry triggers Ca2+ release from peripheral coupling SR that subsequently induces further Ca2+ release from stores in more central regions of the myocyte. Spreading of Ca2+ release from the cell periphery to the centre accounts for [Ca2+]i gradients underlying the whole-cell [Ca2+]i transient. The finding that cat atrial myocytes lack t-tubules demonstrates the functional importance of Ca2+ release from extended junctional (corbular) SR in these cells.


Undiagnosed major risk factors in acute ischaemic stroke patients: frequency, profile, stroke mechanisms and outcome.
There is scarce clinical information about the clinical profile of patients with acute ischaemic stroke with previously undiagnosed major vascular risk factors (UMRFs). This was a retrospective analysis of data from the Acute Stroke Registry and Analysis of Lausanne registry between 2003 and 2018 with univariate and multivariate logistic regression analyses comparing clinical profiles of patients with UMRFs to patients with at least one previously diagnosed MRF (DMRF). In all, 4354 patients (median age 70 years [interquartile range 15.2], 44.7% female) were included after excluding 763 (14.9%) for lack of consent and three for missing information. Amongst 1125 (25.8%) UMRF patients, 69.7% (n = 784) had at least one newly diagnosed MRF and the others none. The newly detected MRFs were dyslipidaemia (61.4%), hypertension (23.7%), atrial fibrillation (10.2%), diabetes mellitus (5.2%), ejection fraction <35% (2.0%) and coronary disease (1.0%). Comparing UMRF patients to DMRF patients, multivariate analysis showed a positive association with lower age, non-Caucasian ethnicity, contraceptive use (<55 years old), smoking (≥55 years old) and patent-foramen-ovale-related stroke mechanism. A negative association was found with pre-stroke antiplatelet use and higher body mass index. Functional outcome did not differ. Cerebrovascular recurrences were similar between groups. In this large single-centre cohort, 69.7% of patients with acute ischaemic stroke and UMRF were newly diagnosed with at least one new MRF, the most common being dyslipidaemia, hypertension or atrial fibrillation. Patients of the UMRF group were younger, more often smokers and on contraceptives, and had more patent-foramen-ovale-related strokes.


[Age-related changes of the hypothalamic-pituitary-adrenal axis in individuals with various types of adaptive behaviour].
The purpose was to study age-related changes of the hypothalamic-pituitary-adrenal (HPA) axis function and their interrelations with features of cognitive aging in individuals with various type of adaptive behaviour on model of laboratory primates. Plasma cortisol (F) and dehydroepiandrosterone sulfate (DHEAS) levels in basal conditions (0900h and 2100h of day) and under acute psycho emotional stress (2-hour moderate restraint) were estimated in blood plasma of 34 young (6-8 years) and 34 old (20-27 years) female rhesus monkeys with various types of adaptive behaviour (aggressive, adequate, depressive-like). We have found that the age-related changes in HPA of monkeys of the depression-like behavior were accompanied by the maximal absolute and relative hypercortisolemia in the basal conditions as well as in the stress conditions. These disturbances in HPA are apparently associated with the higher age-related cognitive disturbances. Moreover, the young aggressive monkeys in comparison with the young monkeys of other behavior groups, demonstrated the higher levels of DHEAS and the minimal molar ratios between F and DHEAS plasma levels. For old animals with the aggressive behavior, these inter-group differences have leveled off. The minimal age-related changes in HPA and the minimal cognitive disturbances have been revealed for the monkeys with the adequate behavior. These results establish that the age-related damage of the HPA axis and the relevant age-related changes in the cognitive processes are the characteristic individual peculiarities of the animals associated with their individual features in the adaptive behavior strategy.


GOrilla: a tool for discovery and visualization of enriched GO terms in ranked gene lists.
Since the inception of the GO annotation project, a variety of tools have been developed that support exploring and searching the GO database. In particular, a variety of tools that perform GO enrichment analysis are currently available. Most of these tools require as input a target set of genes and a background set and seek enrichment in the target set compared to the background set. A few tools also exist that support analyzing ranked lists. The latter typically rely on simulations or on union-bound correction for assigning statistical significance to the results. GOrilla is a web-based application that identifies enriched GO terms in ranked lists of genes, without requiring the user to provide explicit target and background sets. This is particularly useful in many typical cases where genomic data may be naturally represented as a ranked list of genes (e.g. by level of expression or of differential expression). GOrilla employs a flexible threshold statistical approach to discover GO terms that are significantly enriched at the top of a ranked gene list. Building on a complete theoretical characterization of the underlying distribution, called mHG, GOrilla computes an exact p-value for the observed enrichment, taking threshold multiple testing into account without the need for simulations. This enables rigorous statistical analysis of thousand of genes and thousands of GO terms in order of seconds. The output of the enrichment analysis is visualized as a hierarchical structure, providing a clear view of the relations between enriched GO terms. GOrilla is an efficient GO analysis tool with unique features that make a useful addition to the existing repertoire of GO enrichment tools. GOrilla's unique features and advantages over other threshold free enrichment tools include rigorous statistics, fast running time and an effective graphical representation. GOrilla is publicly available at: http://cbl-gorilla.cs.technion.ac.il


Gene expression analysis on biochemical networks using the Potts spin model.
Microarray technology allows us to profile the expression of a large subset or all genes of a cell. Biochemical research over the last three decades has elucidated an increasingly complete image of the metabolic architecture. For less complex organisms, such as Escherichia coli, the biochemical network has been described in much detail. Here, we investigate the clustering of such networks by applying gene expression data that define edge lengths in the network. The Potts spin model is used as a nearest neighbour based clustering algorithm to discover fragmentation of the network in mutants or in biological samples when treated with drugs. As an example, we tested our method with gene expression data from E.coli treated with tryptophan excess, starvation and trpyptophan repressor mutants. We observed fragmentation of the tryptophan biosynthesis pathway, which corresponds well to the commonly known regulatory response of the cells.


High-dose taurine supplementation increases serum paraoxonase and arylesterase activities in experimental hypothyroidism.
1. Hypothyroidism is accompanied by hyperlipidaemia and oxidative stress and is associated with several complications, such as atherosclerosis. Paraoxonase activity has been reported to decrease in several situations associated with atherosclerosis and oxidative stress. In the present study, the effects of different doses of taurine on serum paraoxonase and arylesterase activities, as well as on the serum lipid profile, were investigated in hypothyroid rats. 2. Forty male Sprague-Dawley rats were randomly divided into five groups as follows: Group 1, rats received normal rat chow and tap water; Group 2, rats received standard rat chow + 0.05% propylthiouracil (PTU) in the drinking water; and Groups 3-5, taurine-supplemented PTU groups (standard rat chow + 0.5, 2 or 3% taurine in the drinking water, respectively, in addition to PTU). Paraoxon or phenylacetate were used as substrates to measure paraoxonase and arylesterase activity, respectively. Plasma and tissue malondialdehyde (MDA) levels, indicators of lipid peroxidation, were determined using the thiobarbituric-acid reactive substances method. Serum triglyceride, total cholesterol and high-density lipoprotein-cholesterol (following precipitation with dextran sulphate-magnesium chloride) were determined using enzymatic methods. 3. Serum paraoxonase and arylesterase activities were increased and plasma and tissue MDA levels and serum triglyceride levels were reduced in a dose-dependent manner in taurine-treated hypothyroid rats. Taurine concentrations were positively correlated with enzyme activities and negatively correlated with MDA and triglyceride levels. 4. Further studies are needed to investigate the role of taurine supplementation in hypothyroidism in human subjects.


Effects of larval growth condition and water availability on desiccation resistance and its physiological basis in adult Anopheles gambiae sensu stricto.
Natural populations of the malaria mosquito Anopheles gambiae s.s. are exposed to large seasonal and daily fluctuations in relative humidity and temperature, which makes coping with drought a crucial aspect of their ecology. To better understand natural variation in desiccation resistance in this species, the effects of variation in larval food availability and access to water as an adult on subsequent phenotypic quality and desiccation resistance of adult females of the Mopti chromosomal form were tested experimentally. It was found that, under normal conditions, larval food availability and adult access to water had only small direct effects on female wet mass, dry mass, and water, glycogen and body lipid contents corrected for body size. In contrast, when females subsequently faced a strong desiccation challenge, larval food availability and adult access to water had strong carry-over effects on most measured physiological and metabolic parameters, and affected female survival. Glycogen and water content were the most used physiological reserves in relative terms, but their usage significantly depended on female phenotypic quality. Adult access to water significantly influenced the use of water and body lipid reserves, which subsequently affected desiccation resistance. These results demonstrate the importance of growth conditions and water availability on adult physiological status and subsequent resistance to desiccation.


Measuring delay discounting in a crowdsourced sample: An exploratory study.
Delay discounting is a measure of preferences for smaller immediate rewards over larger delayed rewards. Discounting has been assessed in many ways; these methods have variably and inconsistently involved measures of different lengths (single vs. multiple items), forced-choice methods, self-report methods, online and laboratory assessments, monetary and non-monetary compensation. The majority of these studies have been conducted in laboratory settings. However, over the past 20 years, behavioral data collection has increasingly shifted online. Usually, these experiments involve completing short tasks for small amounts of money, and are thus qualitatively different than experiments in the lab, which are typically more involved and in a strongly controlled environment. The present study aimed to determine how to best measure future discounting in a crowdsourced sample using three discounting measures (a single shot measure, the 27-item Kirby Monetary Choice Questionnaire, and a one-time Matching Task). We examined associations of these measures with theoretically related variables, and assessed influence of payment on responding. Results indicated that correlations between the discounting tasks and conceptually related measures were smaller than in prior laboratory experiments. Moreover, our results suggest providing monetary compensation may attenuate correlations between discounting measures and related variables. These findings suggest that incentivizing discounting measures changes the nature of measurement in these tasks.


[Assessment of severity of the state and intensive care of newborns in the intensive care units].
A point-based evaluation of the severity condition was made on the bases of an analysis of clinical findings (cardiovascular, respiratory, central nervous, urinary and gastrointestinal systems) for 130 newborns who were admitted to the intensive care and resuscitation unit: 3-5 points--severe condition, 6-10 points--highly severe condition and 12-15 points--extremely severe condition. The severity assessment scale as applicable to newborns and based only on the clinical findings enables the practitioner to optimize the conducted therapy in accordance with a severity degree.


Spatial- and time-explicit human damage modeling of ozone depleting substances in life cycle impact assessment.
Depletion of the stratospheric ozone layer is mainly caused by emissions of persistent halocarbons of anthropogenic origin. The resulting increase of solar ultraviolet radiation at the Earth's surface is associated with increased exposure of humans and increased human health damage. Here we assessed the change in human health damage caused by three types of skin cancer and cataract in terms of (healthy) years of life lost per kiloton emission reduction of an ozone-depleting substance (ODS). This so-called characterization factor is used in Life Cycle Assessments (LCAs). Characterization factors are provided for the emissions of five chlorofluorocarbons, three hydrochlorofluorocarbons, three (bromine-containing) halons, carbon tetrachloride, methyl chloroform, and anthropogenic emissions of methyl bromide. We employed dynamic calculations on a global scale for this purpose, taking physical and social geographic data into account such as skin tones, population density, average age, and life expectancy. When emission rates of all ODSs in 2007 are multiplied by our characterization factors, the resulting number of years of life lost may be a factor of 5 higher than reported previously. This increase is merely explained through the global demographic development until 2100 we took into account.


Ammonium persulfate: a safe alternative oxidizing reagent for measuring urinary iodine.
The chloric acid method is most commonly used to obtain accurate and reproducible measurements of iodine and remove interfering substances. Unfortunately chloric acid is a potential hazard, requiring an explosion-proof hood, among other precautions. We have developed a simple, convenient, and economic method for measuring urinary iodine by using 1 mol/L ammonium persulfate, a nonexplosive, nonhazardous chemical, as the oxidizing reagent. The oxidation procedure can be completed in 30 min at a temperature of 91-95 degrees C. The iodine in the urine is then measured by a modification of the traditional colorimetric method of Sandell and Kolthoff. Urine samples (110) collected from a mixed population of healthy males and females, ranging in age from 6 to 79 years and living in the US, were analyzed for urine iodine content by two methods: the proposed ammonium persulfate method and the chloric acid method. The ammonium persulfate method has an intraassay CV of 9.1% at 0.42 +/- 0.04 micromol/L (mean +/- SD), 7.8% at 1.46 +/- 0.11 micromol/L, and 4.0% at 3.54 +/- 0.14 micromol/L. The interassay CV is 10.2% at 0.46 +/- 0.05 micromol/L, and 7.9% at 3.27 +/- 0.26 micromol/L. Recovery of iodine added to urine in vitro was 107%, 94%, and 97% for 0.42 micromol/L, 0.77 micromol/L and 3.64 micromol/L, respectively. The lower limit of detectability was 0.0034 microgram of iodine. Values for iodine in 110 urines measured by the reference chloric acid method ranged from 0.06 to 8.03 micromol/L and by the ammonium persulfate method from 0.05 to 7.4 micromol/L. The persulfate method (y) correlated extremely closely with the reference chloric acid method (x) by the Pearson correlation (y = 0.923x + 0.810 micromol/L, and r = 0.994, Sy/x = 1.841).


Interaction between Vitis vinifera and grape phylloxera: changes in root tissue during nodosity formation.
The interaction between the gall-forming grapevine parasite, phylloxera, and the susceptible grapevine species Vitis vinifera was investigated. Phylloxera and grapevines were cocultivated using both potted and micropropagated grapevines. Development of nodosities on primary roots was studied by microscopy and histochemistry, and nodosities were analysed for biochemical changes and changes in gene expression. Within a nodosity, phylloxera fed at a site in the root cortex. Nodosity development was characterized by swelling of the root tissue distal to the feeding site, lack of development of a suberized endodermis, and starch and amino acid accumulation, and was eventually followed by root necrosis. No evidence of a defence response was observed in pre-necrotic nodosities, but defence-type responses were observed in tissue adjacent to necrotic regions. Changes in gene expression were not detected by northern hybridization using DNA probes encoding a range of V. vinifera transcripts. Nodosities on V. vinifera potentially function as nutrient reservoirs, and defence responses to phylloxera attack were not detected.


Consil bioactive glass particles enhance osteoblast proliferation and selectively modulate cell signaling pathways in vitro.
Consil Bioglass is a commercially available bioactive glass formulation previously shown in clinical studies to support osteogenesis and the repair of bony defects in dogs and cats. Previous in vitro studies confirm that Consil particles are able to bond directly with bone while promoting osteoblast proliferation and extracellular matrix production. However, the cellular mechanisms mediating their clinical effect remain unclear. This study evaluated whether enhancement of osteoblast proliferation by Consil particles is associated with signal transduction. Consil particles maintained the osteoblast phenotype and enhanced proliferation of canine osteoblasts for up to 21 days in culture. Stimulation of proliferation and maintenance of phenotype expression were accompanied by the modulation of selective cell signaling pathways including integrins, the mitogen-activated protein kinases (MAPKs), and the immediate-early gene c-Jun. These genes have been documented to mediate osteoblast growth and differentiation. The signal transduction occurs in a time-dependent manner in which Consil particles induce a decrease in the pattern of MAPK and c-Jun gene transcription from 4 to 24 h and a subsequent return to control levels by 7 days in culture. Our observations suggest that Consil Bioglass particles may provide cues that enhance cell division necessary for facilitating bone regeneration and the repair of bony defects.


Epigenetics targeted protein-vorinostat nanomedicine inducing apoptosis in heterogeneous population of primary acute myeloid leukemia cells including refractory and relapsed cases.
Aberrant epigenetics play a key role in the onset and progression of acute myeloid leukemia (AML). Herein we report in silico modelling based development of a novel, protein-vorinostat nanomedicine exhibiting selective and superior anti-leukemic activity against heterogeneous population of AML patient samples (n=9), including refractory and relapsed cases, and three representative cell lines expressing CD34(+)/CD38(-) stem cell phenotype (KG-1a), promyelocytic phenotype (HL-60) and FLT3-ITD mutation (MV4-11). Nano-vorinostat having ~100nm size exhibited enhanced cellular uptake rendering significantly lower IC50 in AML cell lines and patient samples, and induced enhanced HDAC inhibition, oxidative injury, cell cycle arrest and apoptosis compared to free vorinostat. Most importantly, nanomedicine showed exceptional single-agent activity against the clonogenic proliferative capability of bone marrow derived leukemic progenitors, while remaining non-toxic to healthy bone marrow cells. Collectively, this epigenetics targeted nanomedicine appears to be a promising therapeutic strategy against various French-American-British (FAB) classes of AML. Through the use of a protein-vorinostat agent, exceptional single-agent activity was demonstrated against the clonogenic proliferative capability of bone marrow derived leukemic progenitors, while remaining non-toxic to healthy bone marrow cells. The studied epigenetics targeted nanomedicine approach is a promising therapeutic strategy against various French-American-British classes of acute myeloid leukemia.


Repeated ultraviolet exposure affords the same protection against DNA photodamage and erythema in human skin types II and IV but is associated with faster DNA repair in skin type IV.
We have investigated the photoprotective properties of induced pigmentation using erythema and epidermal DNA photodamage as endpoints. Previously unexposed buttock skin of 12 young, healthy adults (six skin type II and six skin type IV) was exposed daily (Monday to Friday) for 2 wk (days 1-12) with 0.65 minimal erythema dose of solar simulated radiation. Mean skin type IV minimal erythema dose was 1.8-fold greater than for skin type II. Compared to skin type II, solar simulated radiation treatments produced less erythema and more tanning in skin type IV. To assess DNA photodamage, biopsies were taken and prepared for paraffin sections that were stained with a monoclonal antibody for thymine dimers. Thymine dimers were quantified by image analysis. The single exposure data (0.65 and 2 minimal erythema dose) showed that DNA damage was related to physical dose (J per cm2) independent of skin type. Our data also showed that DNA photodamage accumulates in both skin types with repeated, suberythemal doses of solar simulated radiation. On day 12, there were more thymine dimers in skin type IV than skin type II, again indicating that physical rather than biologic dose determines the level of DNA damage. Comparisons on days 12 and 19, however, showed a much greater loss of thymine dimers in skin type IV, suggesting better thymine dimer repair. Protection factors for erythema and thymine dimers were calculated and shown to be about 2 in both skin types. This provides further indirect evidence that DNA is a chromophore for erythema, but also suggests that a tan may not be the major factor in natural photoprotection.


Achieving high-resolution 1H-MRSI of the human brain with compressed-sensing and low-rank reconstruction at 7 Tesla.
Low sensitivity MR techniques such as magnetic resonance spectroscopic imaging (MRSI) greatly benefit from the gain in signal-to-noise provided by ultra-high field MR. High-resolution and whole-slab brain MRSI remains however very challenging due to lengthy acquisition, low signal, lipid contamination and field inhomogeneity. In this study, we propose an acquisition-reconstruction scheme that combines 1H free-induction-decay (FID)-MRSI sequence, short TR acquisition, compressed sensing acceleration and low-rank modeling with total-generalized-variation constraint to achieve metabolite imaging in two and three dimensions at 7 Tesla. The resulting images and volumes reveal highly detailed distributions that are specific to each metabolite and follow the underlying brain anatomy. The MRSI method was validated in a high-resolution phantom containing fine metabolite structures, and in five healthy volunteers. This new application of compressed sensing acceleration paves the way for high-resolution MRSI in clinical setting with acquisition times of 5 min for 2D MRSI at 2.5 mm and of 20 min for 3D MRSI at 3.3 mm isotropic.


Extrinsic allergic alveolitis in the tobacco industry.
A total of 57 subjects who had been exposed to mould dust in the tobacco industry were studied. Their working environment showed exposure to spores of different moulds, and 29 subjects (51%) showed antibodies against one or more of the microbes. Fifteen (26%) had work related respiratory symptoms. Eight (14%) showed slight radiographic pulmonary fibrosis. Spirometry showed a tendency toward restriction and obstruction, especially in small airways. Diffusion capacity was decreased in 18% of the workers. Three clinical cases of typical allergic alveolitis were also found. All this suggests that exposure of spores of different moulds (especially Aspergillus fumigatus) in the manufacture of tobacco products may induce symptoms and signs relating to extrinsic allergic alveolitis.


[Role of mitochondria and reactive oxygen species in the progression of heart failure].
Congestive heart failure (CHF) would be associated with mitochondrial abnormalities and increased of reactive species of oxygen (ROS). To clarify these issues we studied the structure, function of the mitochondrial enzyme nitro oxide synthase inducible (iNOS) and lipoperoxidation of membranes, one of their products through the peroxide nitrite ion (ONOO-), in the heart muscle of patients with heart failure congestive (ICC) grade III and IV (according to New York Heart Association). We included 25 patients who underwent cardiovascular surgery to biopsies of the heart muscle. They were stratified into a group with CHF (n = 18) and control group (n = 7). In di-chas biopsies analyzed the enzymatic activity of mitochondrial complex III spectrophotometrically, which was measured in mM.ubiquinona-1.mg prot, while the mitochondrial morphology was analyzed by the Zeiss electron microscope, the areas were quantified with program Axionvision 4.6. Lipoperoxidation of membranes was measured by the presence of ONOO-by immunohistochemistry against primary antibody against 3-nitrotyrosine was used lab kit system biogenic steptobidin biotin peroxidase (SBA) and coloring triamiobencidina (TAB), it is made with semicuantificacion intensity SCORE test. The statistical test used was ANOVA. The heart muscle of patients with CHF showed that the mitochondrial area was reduced by 78% compared with the control (160.37 μm2 ± 9.87) (936.81 μm2 ± 78.48) p 0.0001. There was also a 70% reduction in complex III activity compared to control (1.9 10-2 mM ubiq.mim-prot 1.mg ± 12.6) (5.79 10-2mM ubiq.mim prot-1.mg ± 36.6) p . The presence of ONOO-was significantly increased in patients with CHF. Alterations ultraestructutural and functional mitochondria found in patients with CHF and increased ROS are involved in the measures of physiopathology CCI and whites should be taken into account for future therapies of this condition.


Effect of 22-oxa-1,25-dihydroxyvitamin D3 on human thyroid cancer cell growth.
To examine whether synthetic vitamin D3 analog, 22-oxa-1,25(OH)2D3 (OCT) has an inhibitory effect on the growth of thyroid carcinoma, we tested the in vitro and in vivo effects of OCT on the growth of a well-differentiated thyroid cancer cell line, NPA. OCT bound to its receptor at the same rate as 1,25(OH)2D3, and inhibited the proliferation of NPA cells in vitro in a dose-dependent manner, similar to that observed with 1,25 (OH)2D3. Northern blot analysis showed that steady-state and fetal bovine serum-stimulated levels of c-myc mRNA were suppressed after 0.5-4 hour treatment with OCT. Transfection studies with the deletion mutants of the 5'-up-stream flanking region of c-myc/chloramphenicol acetyltransferase chimera genes indicated the presence of an OCT responsive element between -410 and -106. Next, we examined OCT effects in implanted NPA tumor cells in nude mice. OCT showed no remarkable hypercalcemic effect compared to 1, 25 (OH2)D3, but OCT and 1, 25 (OH2)D3, had no significant inhibitory effect in vivo after either intra-tumor or intra-peritoneum injection. Our results demonstrate that OCT inhibits the proliferation of well-differentiated thyroid cancer in an in vitro system associated with the suppression of c-myc mRNA, but this inhibitory effect was not reproducible in in vivo model.