The L1 stalk is required for efficient export of nascent large ribosomal subunits in yeast.
The ribosomal protein Rpl1 (uL1 in universal nomenclature) is essential in yeast and constitutes part of the L1 stalk which interacts with E site ligands on the ribosome. Structural studies of nascent pre-60S complexes in yeast have shown that a domain of the Crm1-dependent nuclear export adapter Nmd3, binds in the E site and interacts with Rpl1, inducing closure of the L1 stalk. Based on this observation, we decided to reinvestigate the role of the L1 stalk in nuclear export of pre-60S subunits despite previous work showing that Rpl1-deficient ribosomes are exported from the nucleus and engage in translation. Large cargoes, such as ribosomal subunits, require multiple export factors to facilitate their transport through the nuclear pore complex. Here, we show that pre-60S subunits lacking Rpl1 or truncated for the RNA of the L1 stalk are exported inefficiently. Surprisingly, this is not due to a measurable defect in the recruitment of Nmd3 but appears to result from inefficient recruitment of the Mex67-Mtr2 heterodimer.


Regulation of sarcolemmal Na(+)/H(+) exchanger activity by angiotensin II in adult rat ventricular myocytes: opposing actions via AT(1) versus AT(2) receptors.
Increased sarcolemmal Na(+)/H(+) exchanger activity has been implicated as a mediator of the cardiac actions of angiotensin II. We studied the receptor subtypes and signaling pathways involved in the regulation of sarcolemmal Na(+)/H(+) exchanger activity by angiotensin II in adult rat ventricular myocytes. Cells were loaded with the pH-sensitive fluoroprobe carboxy-seminaphthorhodafluor-1, and acid efflux rates estimated during recovery from intracellular acidosis were used to quantify exchanger activity. Sarcolemmal Na(+)/H(+) exchanger activity was not affected by angiotensin II alone but was increased by angiotensin II plus PD123319 (AT(2) antagonist). In contrast, angiotensin II plus losartan (AT(1) antagonist) or CGP42112A (AT(2) agonist) did not affect exchanger activity. The increase in Na(+)/H(+) exchanger activity induced by angiotensin II plus PD123319 was blocked by losartan, PD98059 (extracellular signal-regulated kinase inhibitor), GF109203X (protein kinase C inhibitor), and tyrphostin AG1478 (epidermal growth factor receptor kinase inhibitor). Extracellular signal-regulated kinase phosphorylation and activity, measured by immunoblot analysis and an immune-complex kinase assay, respectively, were increased significantly by angiotensin II plus PD123319; these increases were blocked by losartan and PD98059. The increase in extracellular signal-regulated kinase phosphorylation induced by angiotensin II plus PD123319 was blocked also by GF109203X and tyrphostin AG1478. These data show that AT(1) stimulation increases sarcolemmal Na(+)/H(+) exchanger activity in adult rat ventricular myocytes and that this response requires extracellular signal-regulated kinase activation through a protein kinase C- and epidermal growth factor receptor-mediated mechanism. The positive effect of AT(1) stimulation on Na(+)/H(+) exchanger activity is counteracted by simultaneous AT(2) stimulation through a mechanism that does not involve direct inhibition of the exchanger or attenuation of extracellular signal-regulated kinase activation.


Open-ended assignments and student responsibility.
An inquiry-based laboratory course was created in an effort to increase student responsibility in learning and to improve teaching in areas related to molecular medicine. Authentic medical cases with both scientific and clinical aspects formed the basis of a project-oriented course that also included student laboratory work focused on the disease-related proteins. Students used basic biochemical techniques to develop and test hypotheses relating their results to the clinical findings. The course also included patient demonstrations to personalize students' knowledge of case presentations, lectures on basic biochemical principles relevant to the molecular basis of the cases, and seminars by invited guests with expertise in translational medicine. Students developed proposals for future research as part of the final examination. An inquiry matrix was used to evaluate the degree of learning responsibility taken during the course. By allowing for openness in how to explore the case including choice of methods and interpretation of unexpected results, students gained confidence in their ability to solve problems, formulate and test hypotheses, and collaborate with both clinical and non-clinical professionals.


Textile-based high-performance hydrogen evolution of low-temperature atomic layer deposition of cobalt sulfide.
Hydrogen is an appealing green energy resource to meet increasing energy demands. To produce hydrogen using the hydrogen evolution reaction (HER), platinum, an expensive and scarce metal, is commonly used and plays a crucial role in maximizing catalytic performance. Transition metal chalcogenides, especially cobalt sulfides (CoSx), are considered an alternative to platinum because of their electrochemical properties, for example, low Tafel slopes and overpotentials. Here, we report a light weight, flexible textile-based HER catalyst through a low-temperature process using the atomic layer deposition (ALD) of CoSx. The electrochemical properties of HER catalysts were investigated and found to be impressive, with a low Tafel slope of 41 mV dec-1 and high exchange current density, demonstrating that these are one of the best characteristics among textile-based HER catalysts. The superb catalytic performances were attributed to the amorphous CoSx phase, confirmed by DFT calculations. This study demonstrates that the integration of HER catalysts with textiles allows the development of highly efficient hydrogen energy production systems.


Absence of vaccinia virus detection in a remote region of the Northern Amazon forests, 2005-2015.
Vaccinia virus (VACV) circulates in Brazil and other South America countries and is responsible for a zoonotic disease that usually affects dairy cattle and humans, causing economic losses and impacting animal and human health. Furthermore, it has been detected in wild areas in the Brazilian Amazon. To better understand the natural history of VACV, we investigated its circulation in wildlife from French Guiana, a remote region in the Northern Amazon forest. ELISA and plaque reduction neutralization tests were performed to detect anti-orthopoxvirus antibodies. Real-time and standard PCR targeting C11R, A56R and A26L were applied to detect VACV DNA in serum, saliva and tissue samples. No evidence of VACV infection was found in any of the samples tested. These findings provide additional information on the VACV epidemiological puzzle. The virus could nevertheless be circulating at low levels that were not detected in areas where no humans or cattle are present.


Expression and subcellular localization of E-cadherin, alpha-catenin, and beta-catenin in 8 feline mammary tumor cell lines.
Protein expression and subcellular localization of E-cadherin, alpha-catenin, and beta-catenin in 8 feline mammary tumor cell lines were examined by western blot analysis and fluorescence immunocytochemistry. A low E-cadherin expression was observed in FNN-m cells. Furthermore, compared to other cell lines, two E-cadherin bands existed in FMC-p1 cells and were localized in the perinuclear region; distinct radial lines were observed in the cytoplasm. A low alpha-catenin expression was observed in FON-m cells, but there were no apparent abnormalities in its localization. In contrast, similar levels of beta-catenin expression and cytoplasmic localization were observed in all cell lines.


Megahertz repetition rate-based lock-in detection scheme for rapid data acquisition in terahertz time domain spectroscopy.
We report a fast pump modulation scheme in a terahertz time domain spectroscopy (THz-TDS) system by utilizing the intensity modulation from the megahertz repetition rate of the pump pulse for lock-in detection. In conventional THz-TDS, the modulation required for the high signal-to-noise ratio lock-in detection is achieved through the use of an optical chopper or an AC bias. Here, we propose the use of an electro-optic modulator (EOM), operated as a pulse picker, to vary the repetition rate of the pump pulse, relative to the probe pulse, allowing us to directly use the megahertz laser repetition rate as the reference modulation frequency for lock-in detection. Our proposed scheme is applicable to all types of pulsed THz emitters, including those that cannot be electronically biased. Since the maximum allowable modulation frequency is limited only by the laser repetition rate and/or by the bandwidth of the EOM, megahertz modulation rates, and, consequently, rapid data acquisition times, become possible. Using our technique, we were able to detect an oscillating signal with frequencies up to 10 kHz, using ∼1 µs integration time per point, ∼100× faster than previously reported values for THz-TDS systems.


Dual Cross-linked Vinyl Vitrimer with Efficient Self-Catalysis Achieving Triple-Shape-Memory Properties.
As an emerging class of dynamic cross-linked network, vitrimers have attracted much attention due to the combination of mechanical advantages of thermosets and recyclability of thermoplastics at an elevated temperature. In particular, most vitrimers with multi-shape memory properties usually involve more than one thermal transition or molecular switch, which might pose a challenge for facile sample fabrication and potentially limits their applications. In pursuit of a more universal and simple route, utilizing commercially available and inexpensive reagents to prepare shape-memory vitrimers with dual cross-linked network from vinyl monomer-derived prepolymers is reported here. Copolymerization of desired vinyl monomers gives prepolymers containing carboxyl and zinc carboxylate groups, which are later converted into vitrimers in a single step by post-curing with diglycidylether of bisphenol A. The Zn2+ ions can not only act as physical crosslinking points through ionic coordination interactions, thus providing the triple-shape-memory properties, but also play the role of catalyst to activate transesterification in the dynamic covalent network. This new self-catalyzed vitrimer has excellent transesterification efficiency, triple-shape-memory properties, and can be sufficiently healed and reprocessed at an elevated temperature. The proposed molecular design of self-catalyzed materials opens a new avenue toward commercially relevant fabrication of high-performance vitrimers with multiple shape-memory properties.


Evolution of a behavior-linked microsatellite-containing element in the 5' flanking region of the primate AVPR1A gene.
The arginine vasopressin V1a receptor (V1aR) modulates social cognition and behavior in a wide variety of species. Variation in a repetitive microsatellite element in the 5' flanking region of the V1aR gene (AVPR1A) in rodents has been associated with variation in brain V1aR expression and in social behavior. In humans, the 5' flanking region of AVPR1A contains a tandem duplication of two approximately 350 bp, microsatellite-containing elements located approximately 3.5 kb upstream of the transcription start site. The first block, referred to as DupA, contains a polymorphic (GT)25 microsatellite; the second block, DupB, has a complex (CT)4-(TT)-(CT)8-(GT)24 polymorphic motif, known as RS3. Polymorphisms in RS3 have been associated with variation in sociobehavioral traits in humans, including autism spectrum disorders. Thus, evolution of these regions may have contributed to variation in social behavior in primates. We examined the structure of these regions in six ape, six monkey, and one prosimian species. Both tandem repeat blocks are present upstream of the AVPR1A coding region in five of the ape species we investigated, while monkeys have only one copy of this region. As in humans, the microsatellites within DupA and DupB are polymorphic in many primate species. Furthermore, both single (lacking DupB) and duplicated alleles (containing both DupA and DupB) are present in chimpanzee (Pan troglodytes) populations with allele frequencies of 0.795 and 0.205 for the single and duplicated alleles, respectively, based on the analysis of 47 wild-caught individuals. Finally, a phylogenetic reconstruction suggests two alternate evolutionary histories for this locus. There is no obvious relationship between the presence of the RS3 duplication and social organization in primates. However, polymorphisms identified in some species may be useful in future genetic association studies. In particular, the presence of both single and duplicated alleles in chimpanzees provides a unique opportunity to assess the functional role of this duplication in contributing to variation in social behavior in primates. While our initial studies show no signs of directional selection on this locus in chimps, pharmacological and genetic association studies support a potential role for this region in influencing V1aR expression and social behavior.


Carnivoran remains from the Malapa hominin site, South Africa.
Recent discoveries at the new hominin-bearing deposits of Malapa, South Africa, have yielded a rich faunal assemblage associated with the newly described hominin taxon Australopithecus sediba. Dating of this deposit using U-Pb and palaeomagnetic methods has provided an age of 1.977 Ma, being one of the most accurately dated, time constrained deposits in the Plio-Pleistocene of southern Africa. To date, 81 carnivoran specimens have been identified at this site including members of the families Canidae, Viverridae, Herpestidae, Hyaenidae and Felidae. Of note is the presence of the extinct taxon Dinofelis cf. D. barlowi that may represent the last appearance date for this species. Extant large carnivores are represented by specimens of leopard (Panthera pardus) and brown hyaena (Parahyaena brunnea). Smaller carnivores are also represented, and include the genera Atilax and Genetta, as well as Vulpes cf. V. chama. Malapa may also represent the first appearance date for Felis nigripes (Black-footed cat). The geochronological age of Malapa and the associated hominin taxa and carnivoran remains provide a window of research into mammalian evolution during a relatively unknown period in South Africa and elsewhere. In particular, the fauna represented at Malapa has the potential to elucidate aspects of the evolution of Dinofelis and may help resolve competing hypotheses about faunal exchange between East and Southern Africa during the late Pliocene or early Pleistocene.


Thermodynamic Genome-Scale Metabolic Modeling of Metallodrug Resistance in Colorectal Cancer.
Mass spectrometry-based metabolomics approaches provide an immense opportunity to enhance our understanding of the mechanisms that underpin the cellular reprogramming of cancers. Accurate comparative metabolic profiling of heterogeneous conditions, however, is still a challenge. Measuring both intracellular and extracellular metabolite concentrations, we constrain four instances of a thermodynamic genome-scale metabolic model of the HCT116 colorectal carcinoma cell line to compare the metabolic flux profiles of cells that are either sensitive or resistant to ruthenium- or platinum-based treatments with BOLD-100/KP1339 and oxaliplatin, respectively. Normalizing according to growth rate and normalizing resistant cells according to their respective sensitive controls, we are able to dissect metabolic responses specific to the drug and to the resistance states. We find the normalization steps to be crucial in the interpretation of the metabolomics data and show that the metabolic reprogramming in resistant cells is limited to a select number of pathways. Here, we elucidate the key importance of normalization steps in the interpretation of metabolomics data, allowing us to uncover drug-specific metabolic reprogramming during acquired metal-drug resistance.


Possible role for platelet-activating factor in neutrophil infiltration in allergic inflammation in rats.
Allergic inflammation was induced by injecting an antigen solution into an air pouch made on the dorsum of immunized rats with the antigen azobenzene-arsonate-conjugated acetyl bovine serum albumin. In this model, leukocyte infiltration into the pouch fluid was prominent 4-8 h after the antigen challenge. Most of the infiltrated leukocytes were neutrophils. Administration of the platelet-activating factor (PAF) antagonists such as CV-3988 and L-652,731 into the air pouch 15 min before and at the time of the antigen challenge failed to suppress leukocyte infiltration at 8 h. However, when the PAF antagonist was injected into an air pouch 4 h after the antigen challenge, neutrophil infiltration at 8 h was suppressed in a dose-dependent manner. Combined treatment with the 5-lipoxygenase inhibitor AA861 and the PAF antagonist did not potentiate the effect of the PAF antagonist, suggesting that participation of leukotriene B4 in neutrophil infiltration in this model is negligible. Eosinophil infiltration was very weak at 8 h, and the PAF antagonist showed no significant effect. At 8 h, the PAF level in the serum of the immunized rats was significantly higher than that of the nonimmunized rats. Intravenous administration of the PAF antagonist 15 min before the antigen challenge suppressed leukocyte infiltration more effectively than local administration into the pouch. These results indicate that PAF plays a significant role in neutrophil infiltration in allergic inflammation.


Effects of acid rain on growth and nutrient concentrations in Scots pine and Norway spruce seedlings grown in a nutrient-rich soil.
The effects of artificially applied acid precipitation on growth and nutrient concentrations of Scots pine (Pinus sylvestris L.) and Norway spruce (Picea abies [L.] Karst.) seedlings were investigated in a long-term acid irrigation experiment in field conditions. Seedlings of northern and southern origin were planted in boxes containing peat and composted soil rich in nutrients, and sprinkler irrigated with water acidified with nitric and sulphuric acids to pH 3 or pH 4 for periods varying from two to three and a half growing seasons during 1986-1989. Water irrigated (pH 5.4-7.6) and non-irrigated groups of seedlings were also included in the experiment. At the end of the experiment needles, main and lateral shoots and roots were collected from the seedlings for the determination of height growth and biomass partitioning, and for the analysis of S, N, Mg, P, K, Ca, Mn and Fe concentrations. The treatment effects compared to the irrigated control were studied using multivariate analyses of variance and covariance. In the pine seedlings the total dry matter production increased by 25-70% compared with the irrigated controls when the total wet deposition to the seedlings exceeded 67 kg S ha(-1) and 36 kg N ha(-1) (e.g. after two growing seasons' exposure of the pH 3 treatment). The increase was mainly due to an increase in needle dry weight (54-72% greater at pH 3) and root weight (20-65% greater at pH 3), whereas the height growth or shoot weight growth were less affected. The northern provenance pine seedlings responded more clearly to the pH 3 irrigation than the southern ones. The treatments had no consistent effects on any of the growth variables studied in the spruce seedlings, however. The pines had higher root and foliage Ca concentrations as a result of the acid irrigation, whereas in spruce, acid rain decreased the Ca concentration in needles and shoots. Root Mn and Fe concentrations were higher in both species as a result of the pH 3 treatment. A higher soil conductivity and Ca concentration resulted from the prolonged pH 3 treatment. The results strongly support the hypothesis that the long-term growth and nutrient allocation response of conifers to acid precipitation is dependent both on the tree species and on the nutritional status of the soil.


Effect of protein binding on the pharmacological activity of highly bound antibiotics.
During antibiotic drug development, media are frequently spiked with either serum/plasma or protein supplements to evaluate the effect of protein binding. Usually, previously reported serum or plasma protein binding values are applied in the analysis. The aim of this study was to evaluate this approach by experimentally measuring free, unbound concentrations for antibiotics with reportedly high protein binding and their corresponding antimicrobial activities in media containing commonly used protein supplements. Free, unbound ceftriaxone and ertapenem concentrations were determined in bacterial growth medium with and without bovine/human serum albumin, as well as adult bovine serum and human plasma using in vitro microdialysis. The corresponding antimicrobial activity was determined in MIC and time-kill curve experiments using Escherichia coli ATCC 25922 and Streptococcus pneumoniae ATCC 6303 as test strains. A semimechanistic maximum effect model was simultaneously fitted to the data and respective EC(50) (concentration at half-maximum effect) values compared. Protein binding differed significantly for ceftriaxone (P < 0.05) between human plasma (76.8 +/- 11.0%) and commercially available bovine (20.2 +/- 8.3%) or human serum albumin (56.9 +/- 16.6%). Similar results were obtained for ertapenem (human plasma, 73.8 +/- 11.6%; bovine serum albumin, 12.4 +/- 4.8%; human serum albumin, 17.8 +/- 11.5%). The MICs and EC(50)s of both strains were significantly increased (P < 0.05) for ceftriaxone when comparing human and bovine serum albumin, whereas the EC(50)s were not significantly different for ertapenem. Free, unbound antibiotic concentrations differed substantially between plasma and protein supplements and correlated well with antimicrobial efficacy. Therefore, free, active concentrations should be measured in the test system instead of correcting for literature protein binding values.


Spinal cord atrophy and disability in multiple sclerosis over four years: application of a reproducible automated technique in monitoring disease progression in a cohort of the interferon beta-1a (Rebif) treatment trial.
Pathology in the cervical spinal cord is considered an important cause of disability in multiple sclerosis. However, the majority of serial studies have failed to find a correlation between spinal cord atrophy and disability. To use a highly reproducible and accurate method to quantify spinal cord area change on three dimensional magnetic resonance imaging and relate this to disability change in patients with multiple sclerosis. 38 patients with multiple sclerosis (20 with the relapsing-remitting (RRMS) form and 18 with the secondary progressive (SPMS) form) were imaged at baseline and at months 6, 12, 18, and 48 during two treatment trials of the high dose subcutaneous thrice weekly interferon beta-1a (IFNbeta, Rebif). Thirty one healthy subjects were also imaged at baseline. Upper cervical cord area (UCCA) was measured using Sobel edge detection. The intraobserver coefficient of variation of the method was 0.42%. A significant reduction in UCCA was detected at month 6 in the placebo group (p = 0.04) and at month 12 for INFbeta (p = 0.03). The mean reduction of UCCA at month 48 was 5.7% for patients initially on placebo who received treatment at 24 months (RRMS) or at 36 months (SPMS), and 4.5% for those on IFNbeta throughout the study (p = 0.35). The change in UCCA was significantly correlated with change in the expanded disability status scale at month 12 (r = 0.4, p = 0.016), month 18 (r = 0.32, p = 0.05), and month 48 (r = 0.4, p = 0.016) in the total cohort. Despite the small number of patients studied and the possible confounding effects of interferon treatment, this study showed that edge detection is reproducible and sensitive to changes in spinal cord area, and that this change is related to changes in clinical disability. This suggests a role for measurement of spinal cord atrophy in monitoring disease progression and possible treatment effects in clinical trails.


Mating, conidiation and pathogenicity of Fusarium graminearum, the main causal agent of the head-blight disease of wheat, are regulated by the MAP kinase gpmk1.
To date, only very little is known about the molecular infection mechanisms of the head-blight pathogen of wheat, Fusarium graminearum (teleomorph Gibberella zeae). Here, we report on the isolation and characterization of the Fus3/Pmk1 mitogen-activated protein kinase homologue Gpmk1 from F. graminearum. Disruption of the gpmk1 gene in F. graminearum results in mutants that are reduced in conidial production, are sexually sterile and are fully apathogenic. This leads to the conclusion that gpmk1 is responsible for signal transduction processes taking place during the most important developmental stages in the life cycle of this fungal pathogen. Thus, Delta gpmk1 mutants are a useful tool to find other important genes involved in plant-infection mechanisms. Previously, only the trichothecene biosynthesis pathway was identified as a virulence factor in F. graminearum. Hence, Gpmk1 is now the second pathogenicity trait to be known in this important plant pathogen.


Derivation of an inhalation reference concentration based upon olfactory neuronal loss in male rats following subchronic acetaldehyde inhalation.
Acetaldehyde inhalation induces neoplastic and nonneoplastic responses in the rodent nasal cavity. This experiment further characterizes the dose-response relationship for nasal pathology, nasal epithelial cell proliferation, and DNA-protein cross-link formation in F-344 rats exposed subchronically to acetaldehyde. Animals underwent whole-body exposure to 0, 50, 150, 500, or 1500 ppm acetaldehyde for 6 h/day, 5 days/wk for up to 65 exposure days. Respiratory tract histopathology was evaluated after 4, 9, 14, 30, and 65 exposure days. Acetaldehyde exposure was not associated with reduced body weight gain or other evidence of systemic toxicity. Histologic evaluation of the nasal cavity showed an increased incidence of olfactory neuronal loss (ONL) following acute to subchronic exposure to > or = 150 ppm acetaldehyde and increased olfactory epithelial cell proliferation following exposure to 1500 ppm acetaldehyde. The severity of the ONL demonstrated dose- and temporal-dependent behaviors, with minimal effects noted at 150-500 ppm acetaldehyde and moderately severe lesions seen in the highest exposure group, with increased lesion severity and extent as the exposure duration increased. Acetaldehyde exposure was also associated with inflammation, hyperplasia, and squamous metaplasia of the respiratory epithelium. These responses were seen in animals exposed to > or = 500 ppm acetaldehyde. Acetaldehyde exposure was not associated with increased DNA-protein cross-link formation in the respiratory or olfactory epithelium. A model of acetaldehyde pharmacokinetics in the nose was used to derive an inhalation reference concentration (RfC) of 0.4 ppm, based on the no-observed-adverse-effect level (NOAEL) of 50 ppm for the nasal pathology seen in this study.


DNA methylation of cancer genome.
DNA methylation plays an important role in regulating normal development and carcinogenesis. Current understanding of the biological roles of DNA methylation is limited to its role in the regulation of gene transcription, genomic imprinting, genomic stability, and X chromosome inactivation. In the past 2 decades, a large number of changes have been identified in cancer epigenomes when compared with normals. These alterations fall into two main categories, namely, hypermethylation of tumor suppressor genes and hypomethylation of oncogenes or heterochromatin, respectively. Aberrant methylation of genes controlling the cell cycle, proliferation, apoptosis, metastasis, drug resistance, and intracellular signaling has been identified in multiple cancer types. Recent advancements in whole-genome analysis of methylome have yielded numerous differentially methylated regions, the functions of which are largely unknown. With the development of high resolution tiling microarrays and high throughput DNA sequencing, more cancer methylomes will be profiled, facilitating the identification of new candidate genes or ncRNAs that are related to oncogenesis, new prognostic markers, and the discovery of new target genes for cancer therapy.


Endoscopic full-thickness resection (eFTR) of colorectal lesions: results from the Dutch colorectal eFTR registry.
Endoscopic full-thickness resection (eFTR) is a minimally invasive resection technique that allows definite diagnosis and treatment for complex colorectal lesions ≤ 30 mm unsuitable for conventional endoscopic resection. This study reports clinical outcomes from the Dutch colorectal eFTR registry. Consecutive patients undergoing eFTR in 20 hospitals were prospectively included. The primary outcome was technical success, defined as macroscopic complete en bloc resection. Secondary outcomes were: clinical success, defined as tumor-free resection margins (R0 resection); full-thickness resection rate; and adverse events. RESULTS : Between July 2015 and October 2018, 367 procedures were included. Indications were difficult polyps (non-lifting sign and/or difficult location; n = 133), primary resection of suspected T1 colorectal cancer (CRC; n = 71), re-resection after incomplete resection of T1 CRC (n = 150), and subepithelial tumors (n = 13). Technical success was achieved in 308 procedures (83.9 %). In 21 procedures (5.7 %), eFTR was not performed because the lesion could not be reached or retracted into the cap. In the remaining 346 procedures, R0 resection was achieved in 285 (82.4 %) and full-thickness resection in 288 (83.2 %). The median diameter of resected specimens was 23 mm. Overall adverse event rate was 9.3 % (n = 34/367): 10 patients (2.7 %) required emergency surgery for five delayed and two immediate perforations and three cases of appendicitis. CONCLUSION : eFTR is an effective and relatively safe en bloc resection technique for complex colorectal lesions with the potential to avoid surgery. Further studies assessing the role of eFTR in early CRC treatment with long-term outcomes are needed.


A Pilot Study of Deep Learning Models for Camera based Hand Hygiene Monitoring in ICU.
Hand hygiene is key to preventing cross-infections in the Intensive Care Unit (ICU). Monitoring of hand washing activities can effectively increase the compliance of clinicians to hand hygiene. In this paper, we explored the feasibility of recognizing clinicians' hand-washing activities using a clinical dataset recorded in ICU using CCTV cameras. We benchmarked three types of hand hygiene detection methods on the dataset with the aim of identifying the 7-step hand washing procedure defined by WHO. Experimental results show that our approach achieves 97% average accuracy for personalized and 67% for generalized modeling. Preliminary results indicate that hand hygiene recognition is subject-dependent, and thus cross-subject modeling or subject-adaptive learning should be used to enhance generalization. The feasibility and challenges of CCTV-camera-based hand hygiene recognition are discussed. The results may contribute to design a hand hygiene scoring and alert system as part of the IoT system in hospitals. The hospital data and code are available at https://github.com/SunnySideUp11/ICU-MH-20.


[About early preclinical diagnosis of gastroduodenal pathology upon exposure to environmental factors].
The paper presents new screening capacities for the early detection of gastroduodenal diseases. The GastroPanel diagnostic system can determine the functional state and indirectly the mucosal structure of different segments of the stomach from blood tests using the enzyme immunoassay. The system permits Helicobacter pylori infection to be detected. Helicobacter pylori is well-known to be the principal cause of the most common gastroduodenal diseases, such as chronic gastritis, including atrophic gastritis regarded as precancer, as well as ulcerative disease of the stomach and duodenum, and distal gastric cancer. The GastroPanel test system is informative, low-informative, and acceptable for the stomach to be evaluated during population-based surveys. It enables gastroduodenal disease to be suspected at the preclinical stage. The GastroPanel test system opens up possibilities for investigating the chronic exposure of the stomach to poor environmental factors over time.


[Role of circulating angiogenic factors in diabetic kidney disease].
This original article contains the authors own data on homeostasis of angiogenic growth factors (vascular endothelial growth factor - VEGF, angiopoietin 1 and 2 - Ang-1, Ang-2) in diabetic kidney disease. The aims of study were evaluation of alteration on serum concentration of circulating VEGF, Ang-l and Ang-2, and of their association with markers of renal damage (albuminuria, glomerular filtration rate) and anemia in patients with diabetes mellitus. We studied 78 patients type I diabetes mellitus (T1DM) and type 2 diabetes mellitus (T2DM). Among this group 37patients had chronic kidney disease. The serum level of VEGF was elevated in T1DM patients and was associated with degree of proteinuria. The serum concentration of Ang-2 was higher in patients with chronic kidney disease (T1DM and T2DM), renal failure (T1DM), proteinuria and anemia (T2DM). Ang-2 strongly associated with albuminuria (T1DM and T2DM), glomerular filtration rate (T1DM) and hemoglobin (T2DM). Obtained results demonstrate that levels of VEGF and Ang-2 (but not Ang-1) are raised in patients with diabetic kidney disease and associated with markers of renal damage and anemia. These data indicate the presence of the disturbance of angiogenic growth factors (VEGF, Ang-2) homeostasis and activity in diabetic patients with chronic kidney disease.


Impact of GP reminders on follow-up of abnormal cervical cytology: a before-after study in Danish general practice.
Dysplasia may progress because of a loss to follow-up after an abnormal cervical cytology. Approximately 18% of Danish women postpone the recommended follow-up, which depends on the cytology results. To investigate if a reminder to the GP about missed follow-up could reduce the proportion of women who fail to act on a recommended follow-up, and to analyse the effect on sociodemographic and general practice variations. A national electronic GP reminder system was launched in Denmark in 2012 to target missed follow-up after screening, opportunistic testing, or surveillance indication. The authors compared follow-up proportions in a national observational before-after study. From national registries, 1.5 million cervical cytologies (from 2009 to 2013) were eligible for inclusion. Approximately 10% had a recommendation for follow-up. The proportion of cervical cytologies without follow-up was calculated at different time points. Results were stratified by follow-up recommendations and sociodemographic characteristics, and changes in practice variation for follow-up were analysed. Fewer women with a recommendation for follow-up missed follow-up 6 months after a GP reminder. Follow-up improved in all investigated sociodemographic groups (age, ethnicity, education, and cohabitation status). Interaction was found for age and cohabitation status. Variation between practices in loss to follow-up was significantly reduced. An electronic GP reminder system showed potential to improve the quality of cervical cancer screening through reduced loss to follow-up.


Determinants of health and wellbeing in refractory epilepsy and surgery: The Patient Reported, ImpleMentation sciEnce (PRIME) model.
This paper offers a new way of understanding the course of a chronic, neurological condition through a comprehensive model of patient-reported determinants of health and wellbeing: The Patient Reported ImpleMentation sciEnce (PRIME) model is the first model of its kind to be based on patient-driven insights for the design and implementation of initiatives that could improve tertiary, primary, and community healthcare services for patients with refractory epilepsy, and has broad implications for other disorders; PRIME focuses on: patient-reported determinants of health and wellbeing, pathways through care, gaps in treatment and other system delays, patient need and expectation, and barriers and facilitators to high-quality care provision; PRIME highlights that in the context of refractory epilepsy, patients value appropriate, clear, and speedy referrals from primary care practitioners and community neurologists to specialist healthcare professionals based in tertiary epilepsy centers. Many patients also want to share in decisions around treatment and care, and gain a greater understanding of their debilitating disease, so as to find ways to self-manage their illness more effectively and plan for the future. Here, PRIME is presented using refractory epilepsy as the exemplar case, while the model remains flexible, suitable for adaptation to other settings, patient populations, and conditions; PRIME comprises six critical levels: 1) The Individual Patient Model; 2) The Patient Relationships Model; 3) The Patient Care Pathways Model; 4) The Patient Transitions Model; 5) The Pre- and Postintervention Model; and 6) The Comprehensive Patient Model. Each level is dealt with in detail, while Levels 5 and 6 are presented in terms of where the gaps lie in our current knowledge, in particular in relation to patients' journeys through healthcare, system intersections, and individuals adaptive behavior following resective surgery, as well as others' views of the disease, such as family members.


Determination of the Critical Speed of a Cracked Shaft from Experimental Data.
In this work, a procedure to obtain an accurate value of the critical speed of a cracked shaft is presented. The method is based on the transversal displacements of the cracked section when the shaft is rotating at submultiples of the critical speed. The SERR (Strain Energy Ralease Rate) theory and the CCL (Crack Closure Line) approach are used to analyse the proposed methodology for considering the behaviour of the crack. In order to obtain the best information and to define the procedure, the orbits and the frequency spectra at different subcritical rotational speed intervals are analyzed by means of the Fast Fourier Transform. The comparison of the maximum values of the FFT peaks within the intervals allows the subcritical speed to be determined, along with the value of the critical speed. When verified, the proposed procedure is applied to shafts with the same geometry and material and with cracks of increasing depth. The results show that the critical speed diminishes with the severity of the crack, as expected. A comparison is made between the critical speed obtained using the vertical and the horizontal displacements, finding no remarkable differences, meaning that in practical applications only one sensor for one of the displacements (in the vertical or horizontal direction) is needed to determine the critical speed. This is one of the main contributions of the paper, as it means that the orbits of the shaft are not needed. Finally, after this study we can conclude that the best results are achieved when the critical speed is obtained using data displacement in only one direction within the intervals around 12 or 13 of the critical speed.


[Progress on pathogenesis and treatment of paraquat-induced pulmonary fibrosis].
Paraquat (PQ) is a highly effective herbicide with contact toxicity. PQ mainly accumulates in the lungs after absorption into the blood circulation. The respiratory function failure caused by PQ-induced lung injury, especially the irreversible pulmonary fibrosis in late phase, is the leading cause of death in patients with PQ poisoning. The mechanism of PQ poisoning is still unclear. Now it is speculated that oxidative stress and inflammation injury are the main pathogenic mechanisms, and abnormal gene expression, mitochondrial damage, loss of pulmonary surfactant, cytokine network and unbalanced matrix metalloproteinases/tissue inhibitors may be also involved in the pathogenesis. In addition to reducing poison absorption and increasing its removal, the current clinical treatment is mainly composed of antioxidant and anti-immune response, but has poor therapeutic effects. Although many novel methods of treatment have been proposed, most of them are still in the experimental stage. It is a hot spot to clarify the mechanism of PQ poisoning and to seek safe and effective treatment of pulmonary fibrosis. This article reviews the research progress on pathogenesis and treatment of PQ-induced pulmonary fibrosis.


Acute cardiorespiratory collapse from heparin: a consequence of heparin-induced thrombocytopenia.
Heparin has rarely been reported to cause acute cardiorespiratory reactions or collapse. Some reports relate this to underlying heparin-induced thrombocytopenia. To confirm and increase awareness of acute life-threatening cardiopulmonary events when patients with heparin-induced thrombocytopenia are re-exposed to heparin. Retrospective observational case series. Four cardiovascular surgery patients were identified in two adjacent large urban hospitals over a 2-yr-period who experienced eight episodes of cardiorespiratory collapse immediately following heparin administration. All had underlying heparin-induced thrombocytopenia. Intravenous boluses of unfractionated heparin were given to four patients with known or previously unrecognized heparin-induced thrombocytopenia. Two patients experienced severe respiratory distress within 15 min for which they required endotracheal intubation. Two other patients experienced cardiac arrest or a lethal arrhythmia within minutes of receiving intravenous heparin. Serologic tests for heparin-induced antibodies were positive in all patients. In three cases, the platelet count was normal or near normal but fell dramatically (71%) immediately following the heparin bolus. Three cases had prior diagnoses of heparin-induced thrombocytopenia, but health care workers administered heparin either unaware of the diagnosis or ignorant of its significance. No patients died, but all required some form of cardiopulmonary resuscitation and subsequent intensive care. Heparin administration to patients with heparin-induced antibodies can result in life-threatening pulmonary or cardiac events. Appreciation of this phenomenon can unmask cases of heparin-induced thrombocytopenia and strengthens the mandate to avoid any heparin exposure in affected patients. Recognition is crucial to avoiding disastrous outcomes.


Direct block of native and cloned (Kir2.1) inward rectifier K+ channels by chloroethylclonidine.
1. We have investigated the inhibition of inwardly rectifying potassium channels by the alpha-adrenergic agonist/antagonist chloroethylclonidine (CEC). We used two preparations; two-electrode voltage-clamp of rat isolated flexor digitorum brevis muscle and whole-cell patch-clamp of cell lines transfected with Kir2.1 (IRK1). 2. In skeletal muscle and at a membrane potential of -50 mV, chloroethylclonidine (CEC), an agonist at alpha2-adrenergic receptors and an antagonist at alpha1x-receptors, was found to inhibit the inward rectifier current with a Ki of 30 microM. 3. The inhibition of skeletal muscle inward rectifier current by CEC was not mimicked by clonidine, adrenaline or noradrenaline and was not sensitive to high concentrations of alpha1-(prazosin) or alpha2-(rauwolscine) antagonists. 4. The degree of current inhibition by CEC was found to vary with the membrane potential (approximately 70% block at -50 mV c.f. approximately 10% block at -190 mV). The kinetics of this voltage dependence were further investigated using recombinant inward rectifier K+ channels (Kir2.1) expressed in the MEL cell line. Using a two pulse protocol, we calculated the time constant for block to be approximately 8 s at 0 mV, and the rate of unblock was described by the relationship tau=exp((Vm+149)/22) s. 5. This block was effective when CEC was applied to either the inside or the outside of patch clamped cells, but ineffective when a polyamine binding site (aspartate 172) was mutated to asparagine. 6. The data suggest that the clonidine-like imidazoline compound, CEC, inhibits inward rectifier K+ channels independently of alpha-receptors by directly blocking the channel pore, possibly at an intracellular polyamine binding site.


Clostridioides difficile SpoVAD and SpoVAE Interact and Are Required for Dipicolinic Acid Uptake into Spores.
Clostridioides difficile spores, like the spores from most endospore-forming organisms, are a metabolically dormant stage of development with a complex structure that conveys considerable resistance to environmental conditions, e.g., wet heat. This resistance is due to the large amount of dipicolinic acid (DPA) that is taken up by the spore core, preventing rotational motion of the core proteins. DPA is synthesized by the mother cell, and its packaging into the spore core is mediated by the products of the spoVA operon, which has a variable number of genes, depending on the organism. C. difficile encodes 3 spoVA orthologues, spoVAC, spoVAD, and spoVAE. Prior work has shown that C. difficile SpoVAC is a mechanosensing protein responsible for DPA release from the spore core upon the initiation of germination. However, the roles of SpoVAD and SpoVAE remain unclear in C. difficile. In this study, we analyzed the roles of SpoVAD and SpoVAE and found that they are essential for DPA uptake into the spore, similar to SpoVAC. Using split luciferase protein interaction assays, we found that these proteins interact, and we propose a model where SpoVAC/SpoVAD/SpoVAE proteins interact at or near the inner spore membrane, and each member of the complex is essential for DPA uptake into the spore core. IMPORTANCE C. difficile spore heat resistance provides an avenue for it to survive the disinfection protocols in hospital and community settings. The spore heat resistance is mainly the consequence of the high DPA content within the spore core. By elucidating the mechanism by which DPA is taken up by the spore core, this study may provide insight into how to disrupt the spore heat resistance with the aim of making the current disinfection protocols more efficient at preventing the spread of C. difficile in the environment.


Injectable dexamethasone-loaded peptide hydrogel for therapy of radiation-induced ototoxicity by regulating the mTOR signaling pathway.
Radiation-induced ototoxicity is associated with inflammation response and excessive reactive oxygen species in the cochlea. However, the effectiveness of many drugs in clinical settings is limited due to anatomical barriers in the inner ear and pharmacokinetic instability. To address this issue, we developed an injectable hydrogel called RADA32-HRN-dexamethasone (RHD). The RHD hydrogel possesses self-anti-inflammatory properties and can self-assemble into nanofibrous structures, ensuring controlled and sustained release of dexamethasone in the local region. Flow cytometry analysis revealed that the uptake of FITC-conjugated RHD gel by hair cells increased in a time-dependent manner. Compared to free dexamethasone solutions, dexamethasone-loaded RHD gel achieved a longer and more controlled release profile of dexamethasone. Additionally, RHD gel effectively protected against the inflammatory response, reduced excessive reactive oxygen species production, and reversed the decline in mitochondrial membrane potentials induced by ionizing radiation, leading to attenuation of apoptosis and DNA damage. Moreover, RHD gel promoted the recovery of outer hair cells and partially restored auditory function in mice exposed to ionizing radiation. These findings validated the protective effects of RHD gel against radiation-induced ototoxicity in both cell cultures and animal models. Furthermore, RHD gel enhanced the activity of the mammalian target of rapamycin (mTOR) signaling pathway, which was inhibited by ionizing radiation, thereby promoting the survival of hair cells. Importantly, intratympanic injections of RHD gel exhibited excellent biosafety and do not interfere with the anti-tumor effects of radiotherapy. In summary, our study demonstrates the therapeutic potential of injectable dexamethasone-loaded RHD hydrogel for the treatment of radiation-induced hearing loss by regulating the mTOR signaling pathway.


Epigenetic MicroRNAs as Prognostic Markers of Postoperative Atrial Fibrillation: A Systematic Review.
Postoperative atrial fibrillation (POAF) is a common complication after cardiac surgery, increasing the risk for adverse outcomes such as perioperative and long-term mortality, stroke, myocardial infarction, and other thromboembolic events. Epigenetic biomarkers show promise as prognostic tools for POAF. Epigenetic changes, such as DNA methylation, histone modification, and microRNAs (miRNA), can result in altered gene expression and the development of various pathological conditions. This systematic review aims to present the current literature on the association between various epigenetic markers and the development of POAF following cardiac surgery. Here, an electronic literature search was performed using MEDLINE, EMBASE, Cochrane Central Register of Controlled Trials, ClinicalTrials.gov, and Google Scholar to identify studies that reported the role of epigenetic markers in the development of POAF. Five of the 6 studies focused on miRNAs and their association with POAF. In POAF patients, the expression of miR-1 and miR-483-5p were upregulated in the right atrial appendage (RAA), while the levels of miR-133A, miR-208a, miR-23a, miR-26a, miR-29a, miR-29b, and miR-29c were decreased in the RAA and venous blood. One study examined cytosines followed by guanines (CpGs) as DNA methylation markers. Across all studies, 488 human subjects who had undergone cardiac surgery were investigated, and 195 subjects (39.9%) developed new-onset POAF. The current literature suggests that miRNAs may play a role in predicting the development of atrial fibrillation after cardiac surgery. However, more robust clinical data are required to justify their role in routine clinical practice.


[Influence of occlusal contacts on tooth displacement].
During function, the tooth displacement path depends on occlusal contacts of the upper and lower teeth. Therefore, we must reconstruct occlusal relation carefully so as not to disturb physiological tooth displacement. In this investigation, tooth displacements were measured with the Displacement Transducer type M-3 in varying occlusal contact. The following conclusions were obtained. 1. The location of occlusal contacts influenced the tooth displacement of the upper first molar. Especially, an occlusal contact on the lingual inclination of the lingual cusp, buccal cusp had great influence. The tooth showed displacement toward the buccal direction, and it differed from the physiological tooth displacement path during clenching at the intercuspal position. 2. The location of occlusal contacts had little influence on the tooth displacement of the lower first molar. The tooth showed displacement toward the lingual direction. It was similar to the physiological tooth displacement. 3. On the upper first molar, the location of occlusal force influenced the Stress-Strain curve. Especially, the occlusal force on the buccal cusp had great influence on the amount of tooth displacement. 4. On the lower first molar, the location of occlusal force influenced the Stress-Strain curve. Especially, the occlusal force on the lingual cusp brought much displacement and a 2-phase displacement path, at times. Therefore, occlusal contacts have close relation to tooth displacement, and any occlusal reconstruction should harmonize with tooth displacement representing periodontal tissue distortion.


A review of studies of heavy users of psychiatric services.
This paper reviews studies of patients who are heavy users of psychiatric services and identifies areas in which further research and evaluation are indicated. Extensive searches were conducted of the English language psychiatric and psychological literature before 1994. Important references from initially identified studies were followed up. More than 200 articles were reviewed, 72 of which are described in this review. The 72 papers were selected because they dealt with three questions: What is heavy service use? What patient characteristics contribute to it? What service delivery characteristics contribute to it? Criteria for identifying and defining heavy users of psychiatric services vary among studies. Few studies of heavy service users have attempted to examine use of all psychiatric services, both inpatient and community based. In most studies, 10 to 30 percent of patients are identified as heavy users, those who utilize between 50 and 80 percent of service resources. This group consists of a constantly changing cohort of patients who generally have psychotic illnesses as well as comorbid personality disorders and high levels of drug and alcohol misuse. Few studies have examined social issues such as isolation, homelessness, and social support, although these factors appear to contribute significantly to heavy service use. Few attempts have been made to define heavy-user groups in fiscal terms. More research on heavy users of psychiatric services is clearly needed to improve providers' ability to plan appropriately targeted mental health services for this disabled group of patients who use expensive resources.


Twenty-seven protein polymorphisms by two-dimensional electrophoresis of serum, erythrocytes, and fibroblasts in two pedigrees.
Twenty-seven independent polymorphic loci were detected by two-dimensional electrophoresis (2DE) of serum, erythrocytes, and fibroblasts in two large families and analyzed for linkage to classical genetic markers. We detected seven serum, four erythrocyte, and 17 fibroblast protein loci that exhibited charge variation in these two families and in a sample of unrelated individuals. The genetic basis of protein variants was confirmed by quantitative gene-dosage dependence and by conformance to Mendelian transmission in the two families, except for four rare variants for which transmission analysis was not possible. Linkage analysis demonstrated that each of the variants represent products of independent loci, with the exception of erythrocyte locus (RBC4), which we also detected in fibroblasts (NC27). Two allozyme polymorphisms, glyoxalase-1 (GLO1) and phosphoglucomutase-3 (PGM3) were specifically identified here based on genotypic concordance and molecular mass. Unknown fibroblast protein (NC22) may be linked to apolipoprotein E (lod score = 2.8 at theta m = theta f = 0), while a serum protein locus (SER1) may be linked to alpha-haptoglobin (lod score = 2.54 at theta m = .20, theta f = .01). Six of seven polymorphic serum loci were previously located on two-dimensional gels: alpha-1 antitrypsin (PI), Gc-globulin (GC), alpha-2 HS glycoprotein (HSGA), alpha-haptoglobin (HP), and two apolipoproteins (APOE and APOA4). Six of 17 polymorphisms detected in fibroblasts were positionally identical to polymorphic loci seen in lymphocytes. These studies indicate a minimum level of average protein charge heterozygosity of approximately 2.2% for the most predominant human cellular proteins and of 5.6% for the most predominant proteins of serum.


Gene space dynamics during the evolution of Aegilops tauschii, Brachypodium distachyon, Oryza sativa, and Sorghum bicolor genomes.
Nine different regions totaling 9.7 Mb of the 4.02 Gb Aegilops tauschii genome were sequenced using the Sanger sequencing technology and compared with orthologous Brachypodium distachyon, Oryza sativa (rice), and Sorghum bicolor (sorghum) genomic sequences. The ancestral gene content in these regions was inferred and used to estimate gene deletion and gene duplication rates along each branch of the phylogenetic tree relating the four species. The total gene number in the extant Ae. tauschii genome was estimated to be 36,371. The gene deletion and gene duplication rates and total gene numbers in the four genomes were used to estimate the total gene number in each node of the phylogenetic tree. The common ancestor of the Brachypodieae and Triticeae lineages was estimated to have had 28,558 genes, and the common ancestor of the Panicoideae, Ehrhartoideae, and Pooideae subfamilies was estimated to have had 27,152 or 28,350 genes, depending on the ancestral gene scenario. Relative to the Brachypodieae and Triticeae common ancestor, the gene number was reduced in B. distachyon by 3,026 genes and increased in Ae. tauschii by 7,813 genes. The sum of gene deletion and gene duplication rates, which reflects the rate of gene synteny loss, was correlated with the rate of structural chromosome rearrangements and was highest in the Ae. tauschii lineage and lowest in the rice lineage. The high rate of gene space evolution in the Ae. tauschii lineage accounts for the fact that, contrary to the expectations, the level of synteny between the phylogenetically more related Ae. tauschii and B. distachyon genomes is similar to the level of synteny between the Ae. tauschii genome and the genomes of the less related rice and sorghum. The ratio of gene duplication to gene deletion rates in these four grass species closely parallels both the total number of genes in a species and the overall genome size. Because the overall genome size is to a large extent a function of the repeated sequence content in a genome, we suggest that the amount and activity of repeated sequences are important factors determining the number of genes in a genome.


Extracerebellar progenitors grafted to the neurogenic milieu of the postnatal rat cerebellum adapt to the host environment but fail to acquire cerebellar identities.
Stem or progenitor cells acquire specific regional identities during early ontogenesis. Nonetheless, there is evidence that cells heterotopically transplanted to neurogenic regions of the developing or mature central nervous system may switch their fate to adopt host-specific phenotypes. Here, we isolated progenitor cells from different germinative sites along the neuraxis where GABAergic interneurons are produced (telencephalic subventricular zone, medial ganglionic eminence, ventral mesencephalon and dorsal spinal cord), and grafted them to the prospective white matter of the postnatal rat cerebellum, at the time when local interneurons are generated. The phenotype acquired by transplanted cells was assessed by different criteria, including expression of region-specific transcription factors, acquisition of morphological and neurochemical traits, and integration in the cerebellar cytoarchitecture. Regardless of their origin, all the different types of donor cells engrafted in the cerebellar parenchyma and developed mature neurons that shared some morphological and neurochemical features with local inhibitory interneurons, particularly in the deep nuclei. Nevertheless, transplanted cells failed to activate cerebellar-specific regulatory genes. In addition, their major structural features, the expression profiles of type-specific markers and the laminar placement in the recipient cortex did not match those of endogenous interneurons generated during the same developmental period. Therefore, although exogenous cells are influenced by the cerebellar milieu and show remarkable capabilities for adapting to the foreign environment, they essentially fail to switch their fate, integrate in the host neurogenic mechanisms and adopt clear-cut cerebellar identities.


Prostate-specific antigen-enhanced testing and risk stratification for chemoprevention trials.
Prostate-specific antigen (PSA) testing could be used to identify men who are at higher future risk of developing clinical prostate cancer or to diagnose prostate cancer earlier in high-risk groups, such as black men or those with a family history of the disease. These cohorts then could be offered chemopreventive clinical trial participation opportunities. The Physicians' Health Study and other longitudinal studies have shown that even between a PSA level of 1.0 and 4.0 ng/mL, the risk of future prostate cancer is incrementally increased. Department of Defense Studies of young men between 15 and 45 show that normal men have very low PSA values. Using a threshold PSA even as low as 1.5 ng/mL for men in their fifth decade is well beyond the 95th percentile of "normal" PSA. Young black men between 40 and 49 years old have a higher risk of prostate cancer than white men and should be pursued for chemoprevention studies. PSA is not perfect. Benign prostatic hyperplasia and inflammation (and, perhaps, other factors) can confound the use of PSA thresholds to identify men for chemoprevention or early detection. Certain chemopreventive agents may affect PSA physiology without affecting the disease process itself creating a meaningless epiphenomenon. Young black men may not generally be receptive to PSA testing or chemopreventive trials.


Changes in nuclear protein during the cell cycle in cultured mast cells separated by zonal centrifugation.
1. Exponentially grown mouse mast cells (cell line P815, strain Y) were separated by zonal centrifugation on a Ficoll gradient. Fractions were allocated to different phases of the cell cycle according to the specific radioactivity of their DNA. 2. Histones were extracted and their thiol content was analysed. The proportion of reduced thiol increased in S phase, decreasing subsequently. 3. The phosphate content of histone F1 and of the other histones reached a peak in early and later S phase respectively. The incorporation of (32)P into these fractions showed a corresponding increase. 4. The timing of histone synthesis was examined. Incorporation of (14)C-labelled amino acids into the histone fractions took place at the same times as phosphorylation. 5. Acid nuclear proteins differ from the histones in incorporating labelled amino acids and (32)P fairly constantly through the cell cycle.


Enhancement of critical current density in a superconducting NbSe2 step junction.
We investigate the transport properties of a NbSe2 nanodevice consisting of a thin region, a thick region and a step junction. The superconducting critical current density of each region of the nanodevice has been studied as a function of temperature and magnetic field. We find that the critical current density has similar values for both the thin and thick regions away from the junction, while the critical current density of the thin region of the junction increases to approximately 1.8 times as compared with the values obtained for the other regions. We attribute such an enhancement of critical current density to the vortex pinning at the surface step. Our study verifies the enhancement of the critical current density by the geometrical-type pinning and sheds light on the application of 2D superconductors.


Thin-slice judgments in the clinical context.
Clinicians make a variety of assessments about their clients, from judging personality traits to making diagnoses, and a variety of methods are available to do so, ranging from observations to structured interviews. A large body of work demonstrates that from a brief glimpse of another's nonverbal behavior, a variety of traits and inner states can be accurately perceived. Additionally, from these "thin slices" of behavior, even future outcomes can be predicted with some accuracy. Certain clinical disorders such as Parkinson's disease and facial paralysis disrupt nonverbal behavior and may impair clinicians' ability to make accurate judgments. In certain contexts, personality disorders, anxiety, depression, and suicide attempts and outcomes can be detected from others' nonverbal behavior. Additionally, thin slices can predict psychological adjustment to divorce, bereavement, sexual abuse, and well-being throughout life. Thus, for certain traits and disorders, judgments from a thin slice could provide a complementary tool for the clinician's toolbox.


Procyanidins from grape seeds protect against phorbol ester-induced oxidative cellular and genotoxic damage.
To evaluate the inhibitory effects of Vitis vinifera procyanidins (PAs) on carcinogen-induced oxidative stress. The single cell gel electrophoresis technique (comet assay) was employed to detect DNA damage induced by the carcinogen phorbol-12-myristate-13-acetate (PMA). The release of hydrogen peroxidase from polymorphonuclear leukocytes (PMNs) was assayed by the horseradish peroxidase-mediated oxidation of phenol red. The microplate assay was used to detect the presence of oxidative products by means of 2',7'-dichlorofluorescin-diacetate (DCFH-DA). The superoxide dismutase (SOD) activity of liver mitochondria was assayed, based on the ability of SOD to inhibit the generation of superoxidate anions by the xanthine-xanthine oxidase system. The malondialdehyde (MDA) level was determined by the thiobarbituric acid (TBA) assay. DNA of NIH3T3 cells was significantly damaged after addition of PMA. The length of the comet tail was observed,while in normal cells the comet tail could not be observed. PAs showed significant protective effects on carcinogen PMA-induced DNA damage. Through assessment of DCFH-DA oxidation, PAs were shown to inhibit the PMA-induced release of hydrogen peroxide by PMNs, and to inhibit respiratory burst activity in NIH3T3 mouse fibroblasts. Ex vivo study showed that serum from rats administered with PAs displayed similar effects in a dose-dependent manner. In addition, PAs suppressed liver mitochondrial lipid peroxidation induced by PMA. PAs protected the activity of SOD and decreased the level of MDA in liver mitochondria damaged by PMA. Dietary PAs from grape seeds protect against carcinogen-induced oxidative cellular and genotoxic damage.


Defective TNF-alpha-induced apoptosis in STAT1-null cells due to low constitutive levels of caspases.
Signal transducers and activators of transcription (STATs) enhance transcription of specific genes in response to cytokines and growth factors. STAT1 is also required for efficient constitutive expression of the caspases Ice, Cpp32, and Ich-1 in human fibroblasts. As a consequence, STAT1-null cells are resistant to apoptosis by tumor necrosis factor alpha (TNF-alpha). Reintroduction of STAT1alpha restored both TNF-alpha-induced apoptosis and the expression of Ice, Cpp32, and Ich-1. Variant STAT1 proteins carrying point mutations that inactivate domains required for STAT dimer formation nevertheless restored protease expression and sensitivity to apoptosis, indicating that the functions of STAT1 required for these activities are different from those that mediate induced gene expression.


MAO-A Inhibitory Potential of Terpene Constituents from Ginger Rhizomes-A Bioactivity Guided Fractionation.
In the search for novel antidepressive drug candidates, bioguided fractionation of nonpolar constituents present in the oleoresin from ginger rhizomes (Zingiber officinale Roscoe, Zingiberaceae) was performed. This particular direction of the research was chosen due to the existing reports on the antidepressive properties of ginger total extract. The search for individual metabolites acting as MAO-A inhibitors, which correspond to the apparent effect of the total extract, is the subject of this work. Hexane extracts from ginger rhizomes were fractionated by using column chromatography (including silica gel impregnated with silver nitrate) and semi-preparative high-performance chromatography. For the activity assessment, an in vitro monoamine oxidase A (MAO-A) inhibition luminescence assay was performed on 10 purified terpenes: 1,8-cineole, α-citronellal, geraniol, β-sesquiphellandrene, γ-terpinen, geranyl acetate, isobornyl acetate, terpinen-4-ol, (E,E)-α-farnesene, and α-zingiberene. Geraniol and (-)-terpinen-4-ol were found to be the strongest enzyme inhibitors with inhibition of 44.1% and 42.5%, respectively, at a concentration of 125 µg/mL. No differences in the inhibition potential were observed for the different groups of terpenes: sesquiterpenes, monoterpenes, or sesquiterpene alcohols; however, the two most active compounds contained a hydroxyl moiety. Terpene constituents from ginger's extract were found to exhibit moderate inhibitory properties against the MAO-A enzyme in in vitro tests.


A Novel Inherently Radiopaque Bead for Transarterial Embolization to Treat Liver Cancer - A Pre-clinical Study.
Embolotherapy using microshperes is currently performed with soluble contrast to aid in visualization. However, administered payload visibility dimishes soon after delivery due to soluble contrast washout, leaving the radiolucent bead's location unknown. The objective of our study was to characterize inherently radiopaque beads (RO Beads) in terms of physicomechanical properties, deliverability and imaging visibility in a rabbit VX2 liver tumor model. RO Beads, which are based on LC Bead® platform, were compared to LC Bead. Bead size (light microscopy), equilibrium water content (EWC), density, X-ray attenuation and iodine distribution (micro-CT), suspension (settling times), deliverability and in vitro penetration were investigated. Fifteen rabbits were embolized with either LC Bead or RO Beads + soluble contrast (iodixanol-320), or RO Beads+dextrose. Appearance was evaluated with fluoroscopy, X-ray single shot, cone-beam CT (CBCT). Both bead types had a similar size distribution. RO Beads had lower EWC (60-72%) and higher density (1.21-1.36 g/cc) with a homogeneous iodine distribution within the bead's interior. RO Beads suspension time was shorter than LC Bead, with durable suspension (>5 min) in 100% iodixanol. RO Beads ≤300 µm were deliverable through a 2.3-Fr microcatheter. Both bead types showed similar penetration. Soluble contrast could identify target and non-target embolization on fluoroscopy during administration. However, the imaging appearance vanished quickly for LC Bead as contrast washed-out. RO Beads+contrast significantly increased visibility on X-ray single shot compared to LC Bead+contrast in target and non-target arteries (P=0.0043). Similarly, RO beads demonstrated better visibility on CBCT in target arteries (P=0.0238) with a trend in non-target arteries (P=0.0519). RO Beads+dextrose were not sufficiently visible to monitor embolization using fluoroscopy. RO Beads provide better conspicuity to determine target and non-target embolization compared to LC Bead which may improve intra-procedural monitoring and post-procedural evaluation of transarterial embolization.


Leg rotation and vastus medialis oblique/vastus lateralis electromyogram activity ratio during closed chain kinetic exercises prescribed for patellofemoral pain.
To examine the effect of leg rotation on the vastus medialis oblique/vastus lateralis (VMO/VL) electromyogram (EMG) activity ratio during two closed chain knee extension exercises prescribed for patellofemoral pain syndrome (PFPS). Subjects performed step-up/step-down (SUSD) and modified wall-slide (MWS), exercises with the leg neutral (SUSD/N, MWS/N), internally rotated (SUSD/I, MWS/I), and externally rotated (SUSD/E, MWS/E), respectively, for determination of root mean square EMG. Six athletically active females symptomatic for PFPS and nine athletically active asymptomatic female control subjects. The EMG activity of the VMO and the VL was collected while the subjects performed the exercises. Root mean square EMG was determined and then normalized to the subjects' maximal voluntary isometric contractions for calculation and analysis of the VMO/VL activity ratio. Leg rotation had no effect on the VMO/VL activity ratio for the symptomatic group. The SUSD/E produced a lower VMO/VL activity ratio compared with the SUSD/N (p < .01) and the SUSD/I (p < .01) in the control group. Similarly, the VMO/VL activity ratio for the MWS/E was lower than for the MWS/I (p < 0.01). These results suggest that external rotation of the leg may affect VMO/VL activity ratio during closed chain knee extension exercises in individuals without PFPS but not in individuals symptomatic for PFPS.


Carcinoma of the bladder T3NxMo treated by preoperative irradiation followed by cystectomy.
Eighty-nine patients with bladder cancer T3NxMo have been treated by preoperative external irradiation (4000 rads in 4 weeks) followed by cystectomy. Five-year survival is 50%. Of patients with T-reduction (T3 becomes P0, P1, or P2) the 5-year survival is 70%. Treatment mortality has been reduced to 3% during the last 2 years. T-reduction cannot be predicted by histologic grade. A second staging by palpation under general anesthesia ("T" 4000) is reliable when T-reduction is felt; however, fibrosis can simulate a persisting T3 mass.


Risk of comorbidities on postoperative outcomes in patients with inflammatory bowel disease.
The effect of comorbidities on postoperative outcomes in patients with inflammatory bowel disease (IBD) has not been explored adequately. We evaluated the prevalence of comorbidities and their effect on postoperative outcomes after an IBD-related operation. The Nationwide Inpatient Sample database was used to identify 35 588 patients with IBD who underwent an IBD-related operation from January 1, 1995, through December 31, 2005. The presence of comorbid illness was assessed using the Elixhauser index. Multiple logistic regression analysis was performed to evaluate the effect of comorbidities on mortality rate after adjusting for age, sex, race, health insurance status, and admission type. Linear regression models were used to evaluate health care resource use. Postoperative mortality was 1.9%. As the number of comorbidities increased (ie, 0, 1, 2, or ≥3), postoperative mortality increased (0.4%, 1.5%, 3.3%, and 7.9%, respectively). Congestive heart failure (odds ratio, 3.50 [95% confidence interval, 2.63-4.62]), liver disease (3.15 [2.00-4.97]), thromboembolic disease (4.19 [3.37-5.21]), and renal disease (8.74 [5.44-14.05]) were associated with a significant increase in mortality rate. Comorbidities associated with an increased risk of mortality also were associated with a significant increase in length of stay and hospital charges. Comorbidities were common in patients with IBD and they significantly increased the risk of postoperative mortality and health care use in patients with IBD.


Different physiological traits underlying increased body fat of fatty (fa/fa) and heterozygous (+/fa) rats.
To find out whether the most characteristic physiological traits distinguishing suckling-age fa/fa pups from lean littermates also differ between +/+ and +/fa littermates, we analyzed the body composition and cold defense of 7- and 16-day-old pups and the plasma concentrations of insulin, glucose, triglycerides, and free fatty acids in 16-day-old pups. Zucker rat x Brown Norway hybrid pups were genotyped by using a molecular marker within 0.5 cM of the fa gene. At both ages the +/fa pups had significantly more body fat than their +/+ littermates. At 7 days this difference was as large as that between +/fa and fa/fa pups, but at 16 days it was only one-seventh of the fa/fa vs. +/fa difference. In contrast, there were no heterozygote differences for three parameters that show crucial abnormalities in the fa/fa pups: thermoregulatory thermogenesis and plasma concentrations of insulin and triglycerides. The physiological mechanisms underlying the increased fat content of +/fa pups thus differ from those known to fuel most of the excessive fat deposition of their fa/fa littermates.


Novel Therapies for Heart Failure - Where Do They Stand?
Despite advances in therapy, patients with heart failure (HF) continue to experience unacceptably high rates of hospitalization and death, as well as poor quality of life. As a consequence, there is an urgent need for new treatments that can improve the clinical course of the growing worldwide population of HF patients. Serelaxin and ularatide, both based on naturally occurring peptides, have potent vasodilatory as well as other effects on the heart and kidneys. For both agents, phase 3 studies that are designed to determine whether they improve outcomes in patients with acute HF have completed enrollment. TRV027, a biased ligand for the type 1 angiotensin receptor with effects that extend beyond traditional angiotensin-receptor blockers is also being studied in the acute HF population. Omecamtiv mecarbil, an inotropic agent that improves myocardial contractility by a novel mechanism, and vericiguat, a drug that stimulates soluble guanylate cyclase, are both being developed to treat patients with chronic HF. Finally, despite the negative results of the CUPID study, gene transfer therapy continues to be explored as a means of improving the function of the failing heart. The basis for the use of these drugs and their current status in clinical trials are discussed. (Circ J 2016; 80: 1882-1891).


13-Hydroxyoctadecadienoic acid is the vessel wall chemorepellant factor, LOX.
We have previously reported that endothelial cells synthesize a cytosol-associated, lipoxygenase-derived metabolite, LOX, which acts as a chemorepellant and, in so doing, maintains the vessel wall thromboresistance. In this study we demonstrate that LOX is a 13-hydroxylinoleic acid (13-OH-18:2) derived from linoleic acid and identical to 13-hydroxy-9-cis,11-trans-octadecadienoic acid, as measured by both reverse phase high pressure liquid chromatography and gas chromatography/mass spectrometry. In addition, we demonstrate that 13-OH-18:2 is produced in significantly greater quantities by endothelial cells than by smooth muscle cels or by fibroblasts. Furthermore, we demonstrate that 13-OH-18:2 is produced in microgram amounts under basal conditions and is decreased by thrombin, calcium ionophore, and trypsin stimulation. And finally, we demonstrate that endothelial cells do not synthesize any significant amounts of lipoxygenase-derived arachidonic acid metabolites either under basal or stimulated conditions unless exogenous arachidonic acid is added. These observations indicate that the major lipoxygenase-derived, chemorepellant metabolite produced by the endothelial is 13-hydroxy-9-cis,11-trans-octadecadienoic acid.


Protective effect of a marine oligopeptide preparation from chum salmon (Oncorhynchus keta) on radiation-induced immune suppression in mice.
A marine oligopeptide preparation (MOP) obtained from Chum Salmon (Oncorhynchus keta) by the method of enzymatic hydrolysis, has been found to enhance the innate and adaptive immunities through stimulation of the secretion of cytokines in mice. The current study aimed to further investigate the protective effect of MOP on radiation-induced immune suppression in mice. Female ICR mice (6-8 weeks old) were randomly divided into three groups, i.e. blank control, irradiation control and MOP (1.350 g kg(-1) body weight) plus irradiation-treated group. MOP significantly increased the survival rate and prolonged the survival times for 30 days after irradiation, and lessened the radiation-induced suppression of T- or B-lymphocyte proliferation, resulting in the recovery of cell-mediated and humoral immune functions. This effect may be produced by augmentation of the relative numbers of radioresistant CD(4) (+) T cells, enhancement of the level of immunostimulatory cytokine, IL-12, reduction of the level of total cellular NF-κB through the induction of IκB in spleen and inhibition of the apoptosis of splenocytes. We propose that MOP be used as an ideal adjuvant therapy to alleviate radiation-induced injuries in cancer patients.


Role of ATF3 as a prognostic biomarker and correlation of ATF3 expression with macrophage infiltration in hepatocellular carcinoma.
The abnormal expression of activating transcription factor 3 (ATF3), a member of the basic leucine zipper (bZIP) family of transcription factors, is associated with carcinogenesis. However, the expression pattern and exact role of ATF3 in the development and progression of hepatocellular carcinoma (HCC) remain unclear. We used UALCAN, ONCOMINE, Kaplan-Meier plotter, and cBioPortal databases to investigate the prognostic value of ATF3 expression in HCC. ATF3 was found to be expressed at low levels in multiple HCC tumor tissues. Moreover, low ATF3 expression was significantly associated with clinical cancer stage and pathological tumor grade in patients with HCC. Therefore, low expression of ATF3 was significantly associated with poor overall survival in patients with HCC. Functional network analysis showed that ATF3 regulates cytokine receptors and signaling pathways via various cancer-related kinases, miRNAs, and transcription factors. ATF3 expression was found to be correlated with macrophage infiltration levels and with macrophage immune marker sets in HCC patients. Using data mining methods, we clarified the role of ATF3 expression and related regulatory networks in HCC, laying a foundation for further functional research. Future research will validate our findings and establish clinical applications of ATF3 in the diagnosis and treatment of HCC.


Comparison of anterior subcutaneous and submuscular transposition of ulnar nerve in treatment of cubital tunnel syndrome: A prospective randomized trial.
This study was designed to compare two methods of surgery, anterior subcutaneous transposition (ASCT) and anterior submuscular transposition (ASMT) of the ulnar nerve in treatment of cubital tunnel syndrome. This randomized trial study was conducted from October 2008 to March 2009 in the Department of Orthopedic Surgery at University Hospital. Forty-eight patients with confirmed cubital tunnel syndrome were randomized in two groups, and each patient received one of two different surgical treatment methods, either ASCT (n = 24) or ASMT (n = 24). In the ASCT technique, the ulnar nerve was transposed and retained in the subcutaneous bed, whereas in the ASMT, the nerve was retained deep in the transected muscular complex, near the median nerve. Patient outcomes, including pain, sensation, muscle strength, and muscle atrophy were compared between groups. The two groups were similar in baseline characteristics. However, those treated with ASMT had a statistically significant reduction in their pain levels compared with ASCT (21 (87.5%) vs 8 (33.3%), P < 0.05). There were no statistically significant differences between the two groups relative to sensation (11 (45.8%) vs 12 (50%)), muscle strength (17 (70.8%) vs 15 (62.5%)), or muscle atrophy (15 (62.5%) vs 17 (70.8%)) (P > 0.05). Our results indicate that ASMT are more efficient than ASCT for managing cubital tunnel syndrome. In patients who had ASMT, there were significant reductions of pain compared with ASCT.


Characterization of a new surface epitope specific for human epithelial cells defined by a monoclonal antibody and application to tumor diagnosis.
In an attempt to characterize the antigens attached to cells of a line established from a human squamous cell carcinoma of the tongue (CAL 27), BALB/c mice were immunized with whole CAL 27 cells; hybridomas were then produced using spleen cells of the animals and cells of an NS1 syngeneic myeloma. A hybridoma secreting a monoclonal antibody was obtained (CALAM 27); CALAM 27 was directed against an epitope attached to the CAL 27 cells. CALAM 27, IgG2a, reacted with a membrane antigen specific to all epithelial cells. After immunoprecipitation, this antigen corresponded to two bands (Mr 22,000 and 54,000). Reactivity disappeared when the tissue was embedded in paraffin but was conserved after fixation with acetone or methanol. This antigen was conserved for both benign and malignant epithelial cell pathologies. The action of CALAM 27 was tested on 80 samples of pleural effusions, ascites, and cerebrospinal fluid samples; after conventional cytological examinations, CALAM 27 failed to recognize either reactive mesothelial cells or meningothelial cells. In addition, the cell structure recognized by CALAM 27 is not found on certain lymphoid tissue cells. CALAM 27 also failed to react with small cell carcinoma of the lung. Its strictly epithelial specificity therefore permits its use for the diagnosis of micrometastases of carcinoma in ascites and cerebrospinal fluid, in pleural effusions, and in bone marrow. CALAM 27 may also prove useful in confirming diagnosis of pathologies suspected to be of epithelial origin.


HBV infection decreases risk of liver metastasis in patients with colorectal cancer: A cohort study.
To evaluate the effect of hepatitis B virus (HBV) infection on liver metastasis of colorectal cancer. A total of 1298 colorectal cancer patients were recruited from January 2001 to March 2005 in this study. Enzyme-linked immunosorbent assay was used to test serum HBV markers for colorectal cancer. Patients were divided into study (infection) group and control (non-infection) group. Clinical features of patients in two groups were compared. Liver metastasis was found in 319 out of the 1298 colorectal cancer patients. The incidence of liver metastasis was significantly lower in study group than in control group (14.2% vs 28.2%, P < 0.01). HBV infection significantly decreased the risk of liver metastasis [hazard ratio (HR): 0.50, 95% confidence interval (95% CI): 0.38-0.66], but the incidence of extrahepatic metastasis was significantly higher in study group than in control group (31.9% vs 17.0%, P < 0.01). The HR was the lowest in chronic hepatitis B group (HR: 0.29, 95% CI: 0.12-0.72). The number of liver metastatic lesions was significantly less in study group than in control group with a higher surgical resection rate. However, no significant difference was found in survival rate between the two groups (P = 0.95). HBV infection decreases the risk of liver metastasis in patients with colorectal cancer and elevates the surgical resection rate of liver metastatic lesions.


Cardiovascular medication: improving adherence.
Adherence to medication is generally defined as the extent to which people take medications as prescribed by their healthcare providers. It can be assessed in many ways (e.g., by self-reporting, pill counting, direct observation, electronic monitoring, or by pharmacy records). This review reports effects of intervention on adherence to cardiovascular medications however adherence has been measured. We conducted a systematic review and aimed to answer the following clinical question: What are the effects of interventions to improve adherence to long-term medication for cardiovascular disease in adults? We searched: Medline, Embase, The Cochrane Library, and other important databases up to April 2010 (Clinical Evidence reviews are updated periodically; please check our website for the most up-to-date version of this review). We included harms alerts from relevant organisations such as the US Food and Drug Administration (FDA) and the UK Medicines and Healthcare products Regulatory Agency (MHRA). We found 39 systematic reviews, RCTs, or observational studies that met our inclusion criteria. We performed a GRADE evaluation of the quality of evidence for interventions. In this systematic review we present information relating to the effectiveness and safety of the following interventions: patient health education, prescriber education, prompting mechanisms, reminder packaging (calendar [blister] packs, multi-dose pill boxes), and simplified dosing.


Development of a TaqMan quantitative reverse transcription PCR assay to detect tilapia lake virus.
Tilapia lake virus disease (TiLVD) is an emerging viral disease associated with high morbidity and mortality in cultured tilapia worldwide. In this study, we have developed and validated a TaqMan quantitative reverse transcription PCR (RT-qPCR) assay for TiLV, targeting a conserved region within segment 10 of the genome. The RT-qPCR assay was efficient (mean ± SD: 96.71 ± 3.20%), sensitive with a limit of detection of 10 RNA viral copies per reaction, and detected TiLV strains from different geographic regions including North America, South America, Africa, and Asia. The intra- and inter-assay variability ranged over 0.18-1.41% and 0.21-2.21%, respectively. The TaqMan RT-qPCR assay did not cross-react with other RNA viruses of fish, including an orthomyxovirus, a betanodavirus, a picornavirus, and a rhabdovirus. Analysis of 91 proven-positive and 185 proven-negative samples yielded a diagnostic sensitivity of 96.7% and a diagnostic specificity of 100%. The TaqMan RT-qPCR assay also detected TiLV RNA in infected Nile tilapia liver tissue extracts following an experimental challenge study, and it successfully detected TiLV RNA in SSN-1 (E-11 clone) cell cultures displaying cytopathic effects following their inoculation with TiLV-infected tissue homogenates. Thus, the validated TaqMan RT-qPCR assay should be useful for both research and diagnostic purposes. Additionally, the TiLV qPCR assay returns the clinically relevant viral load of a sample which can assist health professionals in determining the role of TiLV during disease investigations. This RT-qPCR assay could be integrated into surveillance programs aimed at mitigating the effects of TiLVD on global tilapia production.


The pGRT1 plasmid of Pseudomonas putida DOT-T1E encodes functions relevant for survival under harsh conditions in the environment.
Pseudomonas putida DOT-T1E has the capacity to grow in the presence of high concentrations of toluene. This ability is mainly conferred by an efflux pump encoded in a self-transmissible 133 kb plasmid named pGRT1. Sequence analysis of the pGRT1 plasmid revealed several key features. Most of the genes related to the plasmid maintenance functions show similarity with those encoded on pBVIE04 from Burkholderia vietnamensis G4, and knock-out mutants in several of these genes confirmed their roles. Two additional plasmid DNA fragments were incorporated into the plasmid backbone by recombination and/or transposition; in these DNA regions, apart from multiple recombinases and transposases, several stress-related and environmentally relevant functions are encoded. We report that plasmid pGRT1 not only confers the cells with tolerance to toluene but also resistance to ultraviolet light. We show here the implication of a new protein in solvent tolerance which controls the level of expression of the TtgGHI efflux pump, as well as the implication of a protein with homology to the universal stress protein in solvent tolerance and ultraviolet light resistance. Furthermore, this plasmid encodes functions that allow the cells to chemotactically respond to toluene and participate in iron scavenging.


Climate change and animal diseases in South America.
Climate strongly affects agriculture and livestock production and influences animal diseases, vectors and pathogens, and their habitat. Global warming trends predicted in the 2007 Intergovernmental Panel on Climatic Change (IPCC) report for South America are likely to change the temporal and geographical distribution of infectious diseases, including those that are vector-borne such as bluetongue, West Nile fever, vesicular stomatitis and New World screwworm. Changes in distribution will be partially modulated by El Niño Southern Oscillation events, which will become more frequent and lead to a greater frequency of droughts and floods. Active disease surveillance for animal diseases in South America, particularly for vector-borne diseases, is very poor. Disease reporting is often lacking, which affects knowledge of disease distribution and impact, and preparedness for early response. Improved reporting for animal diseases that may be affected by climate change is needed for better prevention and intervention measures in susceptible livestock, wildlife and vectors in South America. This requires contributions from multidisciplinary experts, including meteorologists, epidemiologists, biologists and ecologists, and from local communities.


Isolation of Yersinia (Pasteurella) pseudotuberculosis from the Palm dove (Streptopelia senegalensis).
Yersinia pseudotuberculosis was isolated from a sick Palm dove (Streptopelia senegalensis) picked up near a turkey farm in Nir-Banim, Israel. Clean birds and a guinea pig inoculated with the isolate died of the infection, or were killed in extremis. Y. pseudotuberculosis showing the same cultural characteristics as the original isolate was recovered from these animals. This is the first reported case of this infection from Israel.


Antagonism of estrogen action with a new benzothiophene derived antiestrogen.
A new benzothiophene derived antiestrogen, LY139481, inhibited the uterotropic action of estradiol in a dose related fashion, and at 1 mg per day suppressed more than 90 percent of estradiol's activity in immature rats. LY139481 induced minimal uterotropic activity, and that activity declined in relation to dose. The relative binding affinity of LY139481 for rat uterine cytosol estrogen receptors was greater than that of estradiol in competitive assays and increased in relation to temperature (2.9 +/- 0.5 x estradiol at 30 degrees C). LY139481 caused estradiol-induced uterine hypertrophy to regress in a manner similar to that which resulted from withdrawal of estradiol treatment. Three successive daily injections of LY139481 slightly increased uterine weight, and blocked additional uterotropic action in response to estradiol and LY139481 administration on subsequent days. Furthermore, ten daily injections of estradiol alone did not increase uterine weight in animals pretreated with LY139481 for three days. In contrast, LY139481 did not prevent the partial uterotropic action of tamoxifen administration.


The Hispanic Americans Baseline Alcohol Survey (HABLAS): predictors of alcohol attitudes and expectancies in Hispanic national groups.
Multiple theoretical frameworks identify attitudes and expectancies as important predictors of alcohol behavior. Few studies have examined demographic predictors of these evaluative and belief-based cognitive mediators in the general population, and none have examined them in large-scale studies of Hispanics, a group at higher risk for drinking behavior and problems. This study probes the extent to which dimensions of attitudes and expectancies share common demographic predictors in a large sample of Puerto Ricans, Cuban-Americans, Mexican-Americans, and South/Central Americans. The 2006 Hispanic Americans Baseline Alcohol Survey (HABLAS) used a multistage cluster sample design to interview 5,224 individuals randomly selected from households in Miami, New York, Philadelphia, Houston, and Los Angeles. This study focused on 2,773 respondents self-identified as current drinkers. Multiple linear regression was used to identify predictors of positive and negative dimensions of attitudes and expectancies, controlling for various background variables. Religious affiliation selectively predicted alcohol attitudes, with Catholics having more positive and fewer negative attitudes than other religious groups. Hispanic group selectively predicted alcohol expectancies, with Cuban-Americans having less positive and less negative expectancies than other groups. Being U.S.-born or male predicted more positive attitudes and expectancies, but birthplace and gender did not predict negative dimensions of attitudes or expectancies. Higher acculturation and more education were linked to a decreased tendency to agree with any item. Age was positively and negatively associated with negative expectancies and positive attitudes, respectively, and having never been married, higher income, and unemployment were each linked to fewer negative attitudes. Although there is some overlap, attitudes and expectancies are influenced by different sociodemographic variables. Positive and negative dimensions of those constructs also show distinct patterns of relations. Prevention and treatment programs targeting cognitive mediators of behavior should be mindful of these differential determinants and future modeling endeavors should incorporate them.


Dietary whole and cracked linseed increases the proportion of oleic and α-linolenic acids in adipose tissues and decreases stearoyl-coenzyme A desaturase, acetyl-coenzyme A carboxylase, and fatty acid synthase gene expression in the longissimus thoracis muscle of Yanbian Yellow cattle.
We hypothesized that supplementation of linseed in a beef cattle fattening diet would increase PUFA concentrations in intramuscular adipose tissue and depress (), (), and () gene expression by decreasing () expression. Conversely, supplemental linseed would upregulate expression of () and () in muscle of Yanbian Yellow steers. Thirty steers were assigned at random to 3 groups of 10 steers fed either the basal diet (corn grain and corn silage-based commercial concentrate [CON]), the CON diet plus 8% whole linseed (WLS; DM basis), or the CON diet plus 8% cracked linseed (CLS; DM basis) for 6 mo. The WLS and CLS supplements did not affect carcass weight, backfat thickness, or marbling scores ( > 0.10) but increased rib eye area and fat color (more yellow; < 0.05). The WLS and CLS diets decreased the proportions of 16:0 and 18:0 and increased the proportions of 18:1-9, 18:3-3, -9, -11 conjugated linoleic acid, total MUFA, and total PUFA in intramuscular, intermuscular, and subcutaneous adipose tissues. The WLS and CLS diets increased and gene expression whereas the supplements depressed , , , and gene expression in longissimus thoracis muscle, relative to CON muscle, consistent with our hypothesis. Because the WLS and CLS treatments did not affect any measure of carcass adiposity, these results indicated that linseed supplements promoted uptake of dietary lipids while concurrently depressing de novo fatty acid biosynthesis in adipose tissue.