Characterization of the optical properties of an infrared blocked impurity band detector.
Si:As blocked impurity band detectors have been partially deprocessed and measured by Fourier transform spectroscopy to determine their transmittance and reflectance at cryogenic temperatures over the wavelength range 2 μm to 40 μm. A method is presented by which the propagation constants can be extracted from an inversion of the transmittance and reflectance data. The effective propagation constants for the active layer from 2 μm to 20 μm were calculated as well as the absorption cross section of arsenic in silicon, which agrees well with previous results from the literature. The infrared absorptance of the full detector was determined, and the analytical method also provides an estimate of absorption in the active layer alone. Infrared absorptance of the active layer is compared to the quantum yield measured by photoelectric means on similar detectors. The optical methods outlined here, in conjunction with standard electronic measurements, could be used to predict the performance of such detectors from measurements of the blanket films from which they are to be fabricated.


Acute renal failure, thrombocytopenia, and elevated liver enzymes after concurrent abuse of alcohol and cocaine.
Cocaine has been associated with known adverse effects on cardiac, cerebrovascular and pulmonary systems. However, the effect of cocaine on other organs has not been extensively reported. A middle age man presented with abdominal pain and nausea after inhalation of crack cocaine. On admission, he was found to be hypertensive and tachycardic. Physical examination revealed mild abdominal tenderness without rebound. Laboratory investigations were significant for acute kidney failure with elevated serum creatinine (3.72 mg/dL), thrombocytopenia (platelet count 74,000/UL), elevated alanine and aspartate transaminases (ALT 331 U/L; AST 462 U/L) and elevated creatine phosphokinase (CPK 5885 U/L). Urine toxicology screening solely revealed cocaine. A clinical diagnosis of cocaine toxicity was made and patient was admitted to the intensive care unit because of multi organ failure. Despite downward trending of liver enzymes during the hospital course, he continued to have residual renal insufficiency and a low platelet count at the time of discharge. In a patient with history of recent cocaine use presenting with these manifestations, cocaine itself should be considered as a likely cause.


[Quantitative limits of the Feulgen reaction: analysis of interference caused by dehistonization and denaturation and renaturation treatments].
The Feulgen reaction intensity (measured with a microdensitometer Vickers M86 on the nucleus of erytrocytes of Xenopus laevis Daud.) is increased after dehistonization according to Brody (1974) only if the dehistonization is made before the fixation in acetic acid. The denaturation and renaturation treatments which should act specifically on the screws of the DNA and therefore should not affect the Feulgen reaction, act in a specific manner, probably going away another histonic components. On the dehistonized material the action of the hydrolysis of the Feulgen reaction would add up to that implicit in the dehistonization treatment and would cause a rapid fall of the values for loss of material as consequence of depolymerization facts, according to Andersson and Kjellstrand (1975). The successive renaturation treatment both on dehistonized and on non dehistonized material does not change significantly the values precedently obtained and this confirms the idea that the rilevability of the Feulgen reaction is not influenced by the treatments "per se" but by the deep "touching" of the chromatin components.


Multimode Kapitza-Dirac interferometry with trapped cold atoms.
We design a multimode interferometer with cold atoms confined in a harmonic trap. A first Kapitza-Dirac pulse creates several spatially addressable modes which are coherently recombined by the harmonic potential and mixed again by a second Kapitza-Dirac pulse. A phase shift among the mode is estimated by fitting the density profile or by measuring the number of atoms in each output mode. The expected sensitivity is rigorously calculated with the Fisher information and the Cramér-Rao lower bound. For the measurement of the gravitational acceleration g we predict, with typical parameters of a compact setup, a temperature independent sensitivity which can exceed, by several orders of magnitude, the sensitivity of current atomic interferometers.


One-step immunoassay for tetrabromobisphenol a using a camelid single domain antibody-alkaline phosphatase fusion protein.
Tetrabromobisphenol A (TBBPA) is a ubiquitous brominated flame retardant, showing widespread environmental and human exposures. A variable domain of the heavy chain antibody (VHH), naturally occurring in camelids, approaches the lower size limit of functional antigen-binding entities. The ease of genetic manipulation makes such VHHs a superior choice to use as an immunoreagent. In this study, a highly selective anti-TBBPA VHH T3-15 fused with alkaline phosphatase (AP) from E. coli was expressed, showing both an integrated TBBPA-binding capacity and enzymatic activity. A one-step immunoassay based on the fusion protein T3-15-AP was developed for TBBPA in 5% dimethyl sulfoxide (DMSO)/phosphate buffered saline (PBS, pH 7.4), with a half-maximum signal inhibition concentration (IC50) of 0.20 ng mL(-1). Compared to the parental VHH T3-15, T3-15-AP was able to bind to a wider variety of coating antigens and the assay sensitivity was slightly improved. Cross-reactivity of T3-15-AP with a set of important brominated analogues was negligible (<0.1%). Although T3-15-AP was susceptible to extreme heat (90 °C), much higher binding stability at ambient temperature was observed in the T3-15-AP-based assay for at least 70 days. A simple pretreatment method of diluting urine samples with DMSO was developed for a one-step assay. The recoveries of TBBPA from urine samples via this one-step assay ranged from 96.7% to 109.9% and correlated well with a high-performance liquid chromatography-tandem mass spectroscopy (HPLC-MS/MS) method. It is expected that the dimerized fusion protein, VHH-AP, will show promising applications in human exposure and environmental monitoring.


Fate of pancreatic nodules induced by raw soya flour in rats.
We have previously shown that rats fed raw soya flour (RSF) for more than four months develop hyperplastic foci of pancreatic acinar cells, which undergo malignant change if feeding RSF is continued throughout the life of the animals. The tendency to undergo malignant change is augmented by the additional use of a genotoxic carcinogen such as azaserine. The present study has sought to examine the reversibility of the focal neoplastic change in the pancreas. Rats fed RSF for 24 weeks and then given a diet not containing soya flour (NSC) had a normal pancreas when killed after 60 weeks of study. When RSF was fed for only 36 weeks, however, some of the rats developed pancreatic cancer even though the diet had been switched to NSC. Similarly, while azaserine in the dose used in the present study does not produce pancreatic cancer in our strain of Wistar rats, coincident administration of RSF for 12 weeks (but not for six weeks) resulted in progression to pancreatic adenoma. Although change from RSF to NSC after 30 weeks resulted in rapid reduction in pancreatic weight and content of RNA, neoplastic foci persisted and became frankly malignant. We conclude that phenotypic reversion to normal of the RSF diet- and azaserine-treated rat pancreas is only possible if RSF alone is fed continuously for not more than about 24 weeks or six weeks if the rats have been exposed to a pancreatic initiating carcinogen.


Calcium metabolism in bullfrog tadpoles (Rana catesbeiana).
Non-feeding bullfrog (Rana catesbeiana) tadpoles can maintain their body Ca concentration in artificial pond waters containing a range of Ca concentrations. The rate of uptake of 45Ca increases when the tadpoles are pre-adapted to a low-Ca pond water and declines when they are pre-adapted to high Ca. Measurement of drinking rate shows that drinking could account for only 5% of the influx. From in vitro experiments, it is estimated that about 70% of the influx occurs accoss the gills, and about 25% through the skin. Calcitonin decreases accumulation of 45Ca while 1,25-(OH)2 vitamin D3 increases it; parathyroid hormone was without effect. Efflux of 45Ca was unchanged by calcitonin or parathyroid hormone.


The NRF2 activator DH404 attenuates adverse ventricular remodeling post-myocardial infarction by modifying redox signalling.
The novel synthetic triterpenoid, bardoxolone methyl, has the ability to upregulate cytoprotective proteins via induction of the nuclear factor erythroid-2-related factor 2 (Nrf2) pathway. This makes it a promising therapeutic agent in disease states characterized by dysregulated oxidative signalling. We have examined the effect of a Nrf2 activator, dihydro-CDDO-trifluoroethyl amide (DH404), a derivative of bardoxolone methyl, on post-infarct cardiac remodeling in rats. DH404, administered from day 2 post myocardial infarction (MI: 30min transient ischemia followed by reperfusion) resulted in almost complete protection against adverse ventricular remodeling as assessed at day 28 (left ventricular end-systolic area: sham 0.14±0.01cm2, MI vehicle 0.29±0.04cm2 vs. MI DH404 0.18±0.02cm2, P<0.05); infarct size (21.3±3.4% MI vehicle vs. 10.9±2.3% MI DH404, P<0.05) with associated benefits on systolic function (fractional shortening: sham 71.9±2.6%, MI vehicle 36.2±1.9% vs. MI DH404 58.6±4.0%, P<0.05). These structural and functional benefits were associated with lower myocardial expression of atrial natriuretic peptide (ANP, P<0.01 vs. MI vehicle), and decreased fibronectin (P<0.01 vs. MI vehicle) in DH404-treated MI rats at 28 days. MI increased glutathionylation of endothelial nitric oxide synthase (eNOS) in vitro - a molecular switch that uncouples the enzyme, increasing superoxide production and decreasing nitric oxide (NO) bioavailability. MI-induced eNOS glutathionylation was substantially ameliorated by DH404. An associated increase in glutaredoxin 1 (Grx1) co-immunoprecipitation with eNOS without a change in expression was mechanistically intriguing. Indeed, in parallel in vitro experiments, silencing of Grx1 abolished the protective effect of DH404 against Angiotensin II-induced eNOS uncoupling. The bardoxolone derivative DH404 significantly attenuated cardiac remodeling post MI, at least in part, by re-coupling of eNOS and increasing the functional interaction of Grx1 with eNOS. This agent may have clinical benefits protecting against post MI cardiomyopathy.


The C-terminal domain LLKIL motif of CXCR2 is required for ligand-mediated polarization of early signals during chemotaxis.
HEK293 cells expressing wild-type CXCR2 recruit PH-Akt-GFP to the leading edge of the cell in response to chemokine. However, in cells expressing mutant CXCR2 defective in AP-2 and HIP binding, i.e. with a mutation in the LLKIL motif, PH-Akt-GFP does not localize to the leading edge in response to ligand. Inhibition of Akt/PKB by transfection of HEK 293 cells with a dominant negative (kinase defective) Akt/PKB inhibits CXCR2 mediated chemotaxis. FRET analysis reveals that membrane-bound activated Cdc42 and Rac1 localize to the leading edge of cells expressing wild-type CXCR2 receptor, but not in cells expressing mutant CXCR2. By contrast, when the activation of Cdc42 and Rac1 are monitored by affinity precipitation assay, cells expressing either wild-type or LLKIL mutant receptors show equivalent ligand induction. Altogether, these data suggest that restricted localized activation of Akt/PKB, Rac1 and Cdc42 is crucial for chemotactic responses and that events mediated by the LLKIL motif are crucial for chemotaxis.


Polymer spacer tunable Purcell-enhanced spontaneous emission in perovskite quantum dots coupled to plasmonic nanowire networks.
Lead halide perovskite quantum dots (PQDs) have recently been proposed as a scalable and color-tunable quantum source, but their slow spontaneous emission creates a mismatch with high-speed nanophotonic devices. Here, we demonstrate fast and bright emission in PQD films coupled to silver nanowire networks (NWKs), in which polyvinyl alcohol (PVA) is used as a spacer to regulate the lossy characteristics of the plasmonic cavity. Compared with bare quartz, the PVA substrate shows a considerable enhancement effect on the apparent emission intensity, but a reduction in the emission rate of PQD excitons. The efficient NWK-PQD coupling generates an increase in the emission intensity of a factor of 6.0 (average 3.4) and simultaneously a 2.4-fold (average 1.9) enhancement in the emission rate. However, an opposite PVA spacer thickness dependence for Purcell factor and quantum yield is observed, indicating that the fast and bright emission would be a trade-off between the Purcell-enhanced radiative rate and large metal loss. These results are believed to provide fundamental guidance on plasmonic cavity design for perovskite-based nanophotonic devices.


Parameter Optimization of Ultrasonic-Microwave Synergistic Extraction of Taxanes from Taxus cuspidata Needles.
Taxanes are the best-known compounds in Taxus cuspidata owing to their strong anticancer effects. However, the traditional taxanes extraction method is the solid-liquid extraction method, which is limited by a large energy consumption and low yield. Therefore, it is urgent to find an efficient method for taxanes extraction. The ultrasonic microwave synergistic extraction (UME) method integrates the cavitation effect of ultrasound and the intensifying heat transfer (ionic conduction and dipole rotation of molecules) effect of microwave to accelerate the release of intracellular compounds and is used in active ingredient extractions. This study aimed to evaluate the performance of UME in extracting taxanes from T. cuspidata needles (dichloromethane-ethanol as extractant). A single-factor experiment, Plackett-Burman design, and the response surface method showed that the optimal UME parameters for taxanes extraction were an ultrasonic power of 300 W, a microwave power of 215 W, and 130 sieve meshes. Under these conditions, the taxanes yield was 570.32 μg/g, which increased by 13.41% and 41.63% compared with the ultrasound (US) and microwave (MW) treatments, respectively. The reasons for the differences in the taxanes yield were revealed by comparing the physicochemical properties of T. cuspidata residues after the UME, US, and MW treatments. The cell structures were significantly damaged after the UME treatment, and numerous tiny holes were observed on the surface. The absorption peaks of cellulose, hemicellulose, and lignin increased significantly in intensity, and the lowest peak temperature (307.40 °C), with a melting enthalpy of -5.19 J/g, was found after the UME treatment compared with the US and MW treatments. These results demonstrate that UME is an effective method (570.32 μg/g) to extract taxanes from T. cuspidata needles by destroying cellular structures.


Pathways from ADHD to early drug use.
This study tests whether attention-deficit hyperactivity disorder (ADHD) increases the risk of early drug use. A community-based sample of 412 low birth weight and 305 normal birth weight children and their mothers initially were assessed when the children were 6 years old with a follow-up assessment at age 11. The relationship of ADHD with drug use varied by level of externalizing problems. Regardless of ADHD status, children with a low level of externalizing problems had a low risk of drug use, and those with the highest level of externalizing problems had a high risk. At the middle level of externalizing problems, ADHD increased the incidence of drug use to the magnitude observed at the high level of externalizing problems, and children with ADHD were at significantly higher risk than those without ADHD (odds ratio = 2.1, p = .03). Findings were similar for low and normal birth weight children. Low parent monitoring and high peer drug use signaled increased risk of drug use for children, independent of ADHD status. Psychostimulant treatment for ADHD was unrelated to risk of drug use. Risk for early drug use in children with ADHD depends on level of associated externalizing problems. Parent monitoring and peer drug use appear to be potential targets for drug prevention for children with ADHD, as well as children in general.


Structure of immunoglobulin gamma 2b heavy chain gene cloned from mouse embryo gene library.
A mouse DNA clone containing the constant part of the immunoglobulin gamma 2b heavy chain was isolated from a mouse gene library. The library was constructed in Charon 4A from a partial EcoRI digest of mouse embryo DNA and was screened with a plasmid (p gamma (11)7) containing a cDNA insert of the heavy chain constant region of the plasmacytoma MPC-11 (1). The Charon 4A clone contains a 14 kb insert which is cleaved by EcoRI into a 6.8 kb and 7.2 kb fragments, of which only the 6.8 kb contains the sequence for gamma 2b heavy chain. Restriction analysis and partial sequence of the insert in p gamma (11) 7 enabled us to obtain three fragments corresponding to the 5' (amino acid 161-302) middle (amino acid 302-443) and 3' (mostly non coding 107 bp) regions of the constant region. Restriction analysis of the Charon 4A clone and hybridisation to these nick translated fragments revealed that the gamma 2b constant region gene contains about 1.5 kb and has three intervening sequences.


Antioxidant properties of probucol released from mesoporous silica.
Antioxidants play a vital role in scavenging reactive oxygen species (ROS) produced by the reduction of molecular oxygen from various cellular mechanisms. Under oxidative stress, an increase in the levels of ROS overwhelms the antioxidant response, causing oxidative damage to biological molecules, and leading to the development of various diseases. Drug compounds with potent antioxidant properties are typically poorly water soluble and highly hydrophobic. An extreme case is Probucol (PB), a potent antioxidant with reported water solubility of 5 ng/ml, and oral bioavailiability of <10%. In this study, PB was loaded in mesoporous silica at various drug loadings to understand the changes to the physical properties of the loaded drug, and it's in vitro drug release. Further in vitro studies were conducted in endothelial and microglia cell models to compare the free radical scavening efficiency of ascorbic acid, PB, and PB release from mesoporous silica particles. Out of the three different mesostructured particles studied, the maximum loading of PB was achieved for large pore mesoporous particles (SBA-15) at 50 wt% drug loading, before complete pore filling was observed. For all materials, loadings above complete pore filling resulted in the recrystallization of PB on the external surface. In vitro drug release measurements showed a rapid dissolution rate at low drug loadings compared to a bimodal release profile of amorphous and crystalline drug at higher drug loadings. PB loaded in mesoporous particle was shown to enhance the antioxidant response to extracellular ROS in the endothelial cell line model, and to intracellular ROS in the microglia cell model. Our results indicate that the antioxidant properties of PB can be significantly improved by using mesoporous silica as a delivery vehicle.


How Progressive Visualizations Affect Exploratory Analysis.
The stated goal for visual data exploration is to operate at a rate that matches the pace of human data analysts, but the ever increasing amount of data has led to a fundamental problem: datasets are often too large to process within interactive time frames. Progressive analytics and visualizations have been proposed as potential solutions to this issue. By processing data incrementally in small chunks, progressive systems provide approximate query answers at interactive speeds that are then refined over time with increasing precision. We study how progressive visualizations affect users in exploratory settings in an experiment where we capture user behavior and knowledge discovery through interaction logs and think-aloud protocols. Our experiment includes three visualization conditions and different simulated dataset sizes. The visualization conditions are: (1) blocking, where results are displayed only after the entire dataset has been processed; (2) instantaneous, a hypothetical condition where results are shown almost immediately; and (3) progressive, where approximate results are displayed quickly and then refined over time. We analyze the data collected in our experiment and observe that users perform equally well with either instantaneous or progressive visualizations in key metrics, such as insight discovery rates and dataset coverage, while blocking visualizations have detrimental effects.


Cholesterol promotes clustering of PI(4,5)P2 driving unconventional secretion of FGF2.
FGF2 is a cell survival factor involved in tumor-induced angiogenesis that is secreted through an unconventional secretory pathway based upon direct protein translocation across the plasma membrane. Here, we demonstrate that both PI(4,5)P2-dependent FGF2 recruitment at the inner plasma membrane leaflet and FGF2 membrane translocation into the extracellular space are positively modulated by cholesterol in living cells. We further revealed cholesterol to enhance FGF2 binding to PI(4,5)P2-containing lipid bilayers. Based on extensive atomistic molecular dynamics (MD) simulations and membrane tension experiments, we proposed cholesterol to modulate FGF2 binding to PI(4,5)P2 by (i) increasing head group visibility of PI(4,5)P2 on the membrane surface, (ii) increasing avidity by cholesterol-induced clustering of PI(4,5)P2 molecules triggering FGF2 oligomerization, and (iii) increasing membrane tension facilitating the formation of lipidic membrane pores. Our findings have general implications for phosphoinositide-dependent protein recruitment to membranes and explain the highly selective targeting of FGF2 toward the plasma membrane, the subcellular site of FGF2 membrane translocation during unconventional secretion of FGF2.


Cytochemical characterization of Triatoma infestans and Panstrongylus megistus salivary gland cells (Hemiptera, Reduviidae, Triatominae).
The Triatominae subfamily has medical sanitary importance, since these insects are vectors of Trypanosoma cruzi, etiologic agent of Chagas Disease, and Trypanosoma rangeli, which develops in the salivary glands and it is frequently found in mixed infections with T. cruzi. Triatomines of Triatoma and Panstrongylus genera possess a salivary gland complex composed of a pair with three well differentiated units: the anterior (D1), median (D2) and posterior (D3). Saliva is secreted during blood meal and antagonizes hemostatic, inflammatory and immunological systems imposed by the host, which facilitate the hematophagy. In order to identify nuclear structures, we studied interphase nuclei of salivary gland cells of adult insects, males and females, of Triatoma infestans and Panstrongylus megistus. The glands were removed from insects, fixed in acetic acid (45%) and lactic acid (50%), squashed and submitted to different cytochemical methods: lacto-acetic orcein, silver ion impregnation, Feulgen reaction, Toluidine Blue, Variant method of critical electrolyte concentration and C-banding. The preparations were examined with a Zeiss Jenaval photomicroscope and photographed. The results evidenced predominance of binucleated cells in D1 and D2 glands and mononucleated ones in D3. In all salivary glands were observed bulky and polyploid nucleus, a clear association between nucleolar and heterochromatic corpuscles, cytoplasmatic metachromasy and many pre-secretion vesicles in cytoplasm. Such characteristics were associated with intense synthesis activity to produce the saliva. Species were mainly differentiated by a larger heterochromatic corpuscle observed only in T. infestans (called as chromocenter), while P. megistus showed a predominance of euchromatin, with some heterochromatic corpuscles just in males. Females of both species showed a smaller quantity of heterochromatin, which could be related to the high metabolism because of the oviposition.


Real-world treatment patterns and outcomes using terlipressin in 203 patients with the hepatorenal syndrome.
Hepatorenal syndrome and acute kidney injury are common complications of decompensated cirrhosis, and terlipressin is recommended as first-line vasoconstrictor therapy. However, data on its use outside of clinical trials are lacking. To assess practice patterns and outcomes around vasoconstrictor use for hepatorenal syndrome in UK hospitals. This was a multicentre chart review study. Data were extracted from medical records of patients diagnosed with hepatorenal syndrome and treated by vasoconstrictor drugs between January 2013 and December 2017 at 26 hospitals in the United Kingdom. The primary outcome was improvement of kidney function, defined as complete response (serum creatinine improved to ≤1.5 mg/dL), partial response (serum creatinine reduction of ≥20% but >1.5 mg/dL) and overall response (complete or partial response). Other outcomes included need for dialysis, mortality, liver transplantation and adverse events. Of the 225 patients included in the analysis, 203 (90%) were treated with terlipressin (median duration, 6 days; range: 2-24 days). Mean (±standard deviation) serum creatinine at vasopressor initiation was 3.25 ± 1.64 mg/dL. Terlipressin overall response rate was 73%. Overall response was higher in patients with mild acute kidney injury (baseline serum creatinine <2.25 mg/dL), compared to those with moderate (serum creatinine ≥2.25 mg/dL and <3.5 mg/dL) or severe (serum creatinine ≥3.5 mg/dL). Ninety-day survival was 86% for all patients (93% for overall responders vs 66% for treatment nonresponders, P < 0.0001). Terlipressin is the most commonly prescribed vasoconstrictor for patients with hepatorenal syndrome in the United Kingdom. Treatment with terlipressin in patients with less severe acute kidney injury (serum creatinine <2.25 mg/dL) was associated with higher treatment responses, and 90-day survival.


Clinical Significance of Spontaneous Pneumomediastinum.
Spontaneous pneumomediastinum (SPM) is classified as free air in the mediastinum in the absence of any precipitating cause. It is relatively uncommon, and the clinical significance and risk associated with SPM is not well understood and has not been widely documented in the literature. Our goals were to determine the outcomes of patients who presented with SPM and to determine predictors of severe pathology associated with SPM. From 2004 through 2013, a retrospective review was conducted of patients who presented with SPM to our institution. Patient demographics, comorbidities, laboratory tests, and esophageal perforation were recorded. In all, 249 patients were discovered to have SPM on chest radiograph or computed tomography scan. Mean age was 38.7 years (range, 17 to 81). Sixty-one percent of patients (151 of 249) were male. Ten percent of all patients (24 of 249) were ultimately discovered to have esophageal perforation, determined by upper endoscopy, upper gastrointestinal series, or intraoperatively during emergent surgery. Age (p < 0.01), pleural effusion (p < 0.01), and elevated white blood cell count (p < 0.01) were the only significant risk factors for esophageal perforation on multivariate analysis. Spontaneous pneumomediastinum is usually associated with a benign clinical course. Risk factors for esophageal perforation in these patients include age, elevated white blood cell count, and a pleural effusion. In the absence of abnormal laboratory values or associated radiologic findings, the majority of patients with SPM may be safely observed without the need for further diagnostic testing.


Up-regulation of glycohydrolases in Alzheimer's Disease fibroblasts correlates with Ras activation.
The lysosomal system is up-regulated in the brain of patients with Alzheimer's Disease (AD), as demonstrated by previous experiments carried out in postmortem samples of brain patients. In this paper we provide evidence that an up-regulation of lysosomal glycohydrolases (alpha-D-mannosidase, beta-D-hexosaminidase, and beta-D-galactosidase) takes place in skin fibroblasts from AD patients affected either by sporadic or familial forms and is detectable also in presymptomatic subjects carrying the above mutations but healthy at the time of skin biopsy. This increase of enzyme activity is consequent to a transcriptional up-regulation. The oncogene Ras appears to be involved in the regulation of enzymatic activity. A parallel increase of Ras transcript and Ras protein, without an increase of p44/p42 MAPK activation was revealed in the same AD fibroblasts. An activation of p38 MAPK already described to occur in neurodegenerative diseases such as Alzheimer's, was also found in fibroblasts derived from AD patients. High levels of expression of the constitutively active form of Ras in normal or AD fibroblasts induced glycohydrolases up-regulation. Overall results demonstrated that glycohydrolases up-regulation, as well as Ras up-regulation, are early markers of AD, detectable at peripheral level, and good candidates to be exploited for diagnostic purposes. These data also provide the first proof for a role of Ras in regulating lysosomal glycohydrolases expression.


Effect of prostaglandin E1 and steroid on healing colonic anastomoses.
The effect of prostaglandin E1 (PGE1) and corticosteroids alone and in combination were studied in the healing rat colon to determine whether PGE1 could not only improve healing but reverse the negative effect of steroids on colonic wound healing. Colonic anastomoses were performed in 144 male Sprague-Dawley rats divided into four groups. The control group (I) received no further treatment. The steroid group (II) received cortisone acetate (5 mg/kg/day) beginning six days preoperatively and continuing until sacrifice. The PGE1 group (III) received 2 micrograms of PGE1 intra-aortically at surgery and for three days postoperatively. The combination PGE1/steroid group (IV) received both drugs in the same doses as those in Groups II and III. Animals were sacrificed on postoperative days 6, 10, and 14. Wound healing was evaluated by hydroxyproline content, bursting pressures, and histology. The hydroxyproline assay at day 10 revealed that steroid-treated rats have significantly lower levels than any other group. The PGE1 group (III) had the highest level of significance in comparison to the steroid group (II) (P = 0.001). The addition of PGE1 to steroid (Group IV) appeared to abolish the negative effect of the steroid as measured by hydroxyproline content on day 10 (P = 0.038). When measuring bursting pressures, the PGE1 group (III) had significantly higher pressures than any other group at day 10. However, no amelioration of the steroid effect on bursting pressures was seen. Histologic evaluation of the anastomosis did not reveal any significant differences among the four groups. PGE1 reverses the negative effect of the steroid on hydroxyproline levels at day 10. Furthermore, using bursting pressure as a parameter of wound healing, administration of PGE1 results in significantly improved anastomotic healing at day 10.


Global analysis of RNA secondary structure in two metazoans.
The secondary structure of RNA is necessary for its maturation, regulation, processing, and function. However, the global influence of RNA folding in eukaryotes is still unclear. Here, we use a high-throughput, sequencing-based, structure-mapping approach to identify the paired (double-stranded RNA [dsRNA]) and unpaired (single-stranded RNA [ssRNA]) components of the Drosophila melanogaster and Caenorhabditis elegans transcriptomes, which allows us to identify conserved features of RNA secondary structure in metazoans. From this analysis, we find that ssRNAs and dsRNAs are significantly correlated with specific epigenetic modifications. Additionally, we find key structural patterns across protein-coding transcripts that indicate that RNA folding demarcates regions of protein translation and likely affects microRNA-mediated regulation of mRNAs in animals. Finally, we identify and characterize 546 mRNAs whose folding pattern is significantly correlated between these metazoans, suggesting that their structure has some function. Overall, our findings provide a global assessment of RNA folding in animals.


Holocene sea ice variability driven by wind and polynya efficiency in the Ross Sea.
The causes of the recent increase in Antarctic sea ice extent, characterised by large regional contrasts and decadal variations, remain unclear. In the Ross Sea, where such a sea ice increase is reported, 50% of the sea ice is produced within wind-sustained latent-heat polynyas. Combining information from marine diatom records and sea salt sodium and water isotope ice core records, we here document contrasting patterns in sea ice variations between coastal and open sea areas in Western Ross Sea over the current interglacial period. Since about 3600 years before present, an increase in the efficiency of regional latent-heat polynyas resulted in more coastal sea ice, while sea ice extent decreased overall. These past changes coincide with remarkable optima or minima in the abundances of penguins, silverfish and seal remains, confirming the high sensitivity of marine ecosystems to environmental and especially coastal sea ice conditions.


The importance of intramolecular conductivity in three dimensional molecular solids.
Recent years have seen tremendous progress towards understanding the relation between the molecular structure and function of organic field effect transistors. The metrics for organic field effect transistors, which are characterized by mobility and the on/off ratio, are known to be enhanced when the intermolecular interaction is strong and the intramolecular reorganization energy is low. While these requirements are adequate when describing organic field effect transistors with simple and planar aromatic molecular components, they are insufficient for complex building blocks, which have the potential to localize a carrier on the molecule. Here, we show that intramolecular conductivity can play a role in controlling device characteristics of organic field effect transistors made with macrocycle building blocks. We use two isomeric macrocyclic semiconductors that consist of perylene diimides linked with bithiophenes and find that the trans-linked macrocycle has a higher mobility than the cis-based device. Through a combination of single molecule junction conductance measurements of the components of the macrocycles, control experiments with acyclic counterparts to the macrocycles, and analyses of each of the materials using spectroscopy, electrochemistry, and density functional theory, we attribute the difference in electron mobility of the OFETs created with the two isomers to the difference in intramolecular conductivity of the two macrocycles.


Hereditary hydrocephalus internus in a laboratory strain of golden hamsters ( Mesocricetus auratus).
Golden hamsters of one common laboratory strain had a high incidence of hydrocephalus internus. When a severity score of hydrocephalus was used, a major autosomal recessive locus could be identified. However, when a binary score (hydrocephalus, no hydrocephalus) was used, no such major locus could be detected and results of test matings were not consistent with Mendelian inheritance. Golden hamsters with severe forms of hydrocephalus had a dorsally compressed and ventrally intact hippocampus. Implications for the behavior and well-being of affected hamsters are unknown but researchers using this strain should be aware of the likely presence of hydrocephalus.


Changes in Diversity and Composition of Rhizosphere Bacterial and Fungal Community between Resistant and Susceptible Pakchoi under Plasmodiophora brassicae.
Plasmodiophora brassicae (P. brassicae) is a soil-born pathogen worldwide and can infect most cruciferous plants, which causes great yield decline and economic losses. It is not well known how microbial diversity and community composition change during P. brassicae infecting plant roots. Here, we employed a resistant and a susceptible pakchoi cultivar with and without inoculation with P. brassicae to analyze bacterial and fungal diversity using 16S rRNA V3-V4 and ITS_V1 regions, respectively. 16S rRNA V3-V4 and ITS_V1 regions were amplified and sequenced separately. Results revealed that both fungal and bacterial diversity increased, and composition was changed in the rhizosphere soil of the susceptible pakchoi compared with the resistant cultivar. In the four groups of R_mock, S_mock, R_10d, and S_10d, the most relatively abundant bacterium and fungus was Proteobacteria, accounting for 61.92%, 58.17%, 48.64%, and 50.00%, respectively, and Ascomycota, accounting for 75.11%, 63.69%, 72.10%, and 90.31%, respectively. A total of 9488 and 11,914 bacteria were observed uniquely in the rhizosphere soil of resistant and susceptible pakchoi, respectively, while only 80 and 103 fungi were observed uniquely in the correlated soil. LefSe analysis showed that 107 and 49 differentially abundant taxa were observed in bacteria and fungi. Overall, we concluded that different pakchoi cultivars affect microbial diversity and community composition, and microorganisms prefer to gather around the rhizosphere of susceptible pakchoi. These findings provide a new insight into plant-microorganism interactions.


Sulfamate-Tethered Aza-Wacker Strategy for a Kasugamine Synthon.
We present our preparation of a kasugamine synthon, which proceeds in 14 steps from a literature epoxide. We expect that this kasugamine derivative can be used for the total syntheses of kasugamycin, minosaminomycin, and analogue antibiotics. A key step in the synthesis is our laboratory's sulfamate-tethered aza-Wacker cyclization.


Signs of current suicidality in men: A systematic review.
Suicide signs have been identified by expert consensus and are relied on by service providers, community helpers' and family members to identify suicidal men. Whether signs that are reported in suicide literature accurately describe male presentations of suicidality is unclear. A systematic review of the literature was conducted to identify male-specific signs of current suicidality and identify gaps in the literature for future research. Searches through Medline, CINAHL, PsychInfo and the Behavioral Sciences Collection, guided by the PRISMA-P statement, identified 12 studies that met the study eligibility criteria. Although the results generally reflected suicide signs identified by expert consensus, there is little research that has examined male-specific signs of the current suicidal state. This review highlights the need for scientific research to clarify male presentation of suicidality. Implications for future research to improve the prompt identification of suicidal men are discussed.


Deep-Learning-Assisted Stratification of Amyloid Beta Mutants Using Drying Droplet Patterns.
The development of simple and accurate methods to predict mutations in proteins remains an unsolved challenge in modern biochemistry. It is discovered that critical information about primary and secondary peptide structures can be inferred from the stains left behind by their drying droplets. To analyze the complex stain patterns, deep-learning neuronal networks are challenged with polarized light microscopy images derived from the drying droplet deposits of a range of amyloid beta (1-42) (Aβ42 ) peptides. These peptides differ in a single amino acid residue and represent hereditary mutants of Alzheimer's disease. Stain patterns are not only reproducible but also result in comprehensive stratification of eight amyloid beta (Aβ) variants with predictive accuracies above 99%. Similarly, peptide stains of a range of distinct Aβ42 peptide conformations are identified with accuracies above 99%. The results suggest that a method as simple as drying a droplet of a peptide solution onto a solid surface may serve as an indicator of minute, yet structurally meaningful differences in peptides' primary and secondary structures. Scalable and accurate detection schemes for stratification of conformational and structural protein alterations are critically needed to unravel pathological signatures in many human diseases such as Alzheimer's and Parkinson's disease.


Arbitrary structuring of two-dimensional photonic crystals by use of phase-only Fourier gratings.
The computer-generated holography technique is applied to the structuring of two-dimensional (2D) photonic crystals with inherently embedded arbitrary defects. The technique uses phase-only Fourier gratings as a generator of spot arrays in the focal plane, such that a single exposure produces a 2D array of focused spots with desired defects or modifications in the lattice structure. We demonstrate several types of large-area 2D lattice structures with square, hexagonal, or hybrid lattices embedded with point and (or) line defects. Scanning the Fourier plane in the depth direction throughout multiphoton polymerization media allows 3D lattices with stacked defect layers to be formed.


Recommendations to Streamline and Standardize Clinical Trial Site Feasibility Assessments: An ASCO Research Statement.
Feasibility assessments (FAs) are important to establish site capabilities to conduct clinical trials and their suitability for specific trials. However, current FA methods used by biotechnology and pharmaceutical (biotech-pharma) trial sponsors and contract research organizations (CROs) are costly, inefficient, unnecessarily burdensome, and resource intensive. These methods delay trial start-up, act as a barrier to site participation, and ultimately reduce timely patient access to clinical trials and novel treatments. An ASCO Task Force was convened to assess the specific burdens and challenges with FAs and to develop recommendations to improve their efficiencies and effectiveness. Stakeholders (including trial sites, biotech-pharma sponsors, and CROs) provided insights into challenges and offered solutions through two surveys and an in-person meeting. The Task Force used the feedback to formulate consensus recommendations to improve FAs for oncology clinical trials. Three key recommendations were identified for application across all biotech-pharma sponsored trials: (1) implement a streamlined and uniform FA process across trials and sponsors; (2) minimize and standardize questions; and (3) leverage technology to centralize FAs, facilitate communications, and reduce redundancies. There is an urgency to improve the current FA process, which is costly, inconsistent, inefficient, labor intensive, and of uncertain effectiveness. All stakeholders stand to benefit from implementing these recommendations, which aim to minimize burdens and ensure that more trial sites and patients have timely access to oncology clinical trials. To have meaningful impact, adoption and consistent execution of these recommendations across all trials, sponsors, CROs, and sites are essential.


Knowledge-Based Strategy to Improve Ligand Pose Prediction Accuracy for Lead Optimization.
Accurately predicting how a small molecule binds to its target protein is an essential requirement for structure-based drug design (SBDD) efforts. In structurally enabled medicinal chemistry programs, binding pose prediction is often applied to ligands after a related compound's crystal structure bound to the target protein has been solved. In this article, we present an automated pose prediction protocol that makes extensive use of existing X-ray ligand information. It uses spatial restraints during docking based on maximum common substructure (MCS) overlap between candidate molecule and existing X-ray coordinates of the related compound. For a validation data set of 8784 docking runs, our protocol's pose prediction accuracy (80-82%) is almost two times higher than that of one unbiased docking method software (43%). To demonstrate the utility of this protocol in a project setting, we show its application in a chronological manner for a number of internal drug discovery efforts. The accuracy and applicability of this algorithm (>70% of cases) to medicinal chemistry efforts make this the approach of choice for pose prediction in lead optimization programs.


H-DNA and Z-DNA in the mouse c-Ki-ras promoter.
The mouse c-Ki-ras protooncogene promoter contains a homopurine-homopyrimidine domain that exhibits S1 nuclease sensitivity in vitro. We have studied the structure of this DNA region in a supercoiled state using a number of chemical probes for non-B DNA conformations including diethyl pyrocarbonate, osmium tetroxide, chloroacetaldehyde, and dimethyl sulfate. The results demonstrate that two types of unusual DNA structures formed under different environmental conditions. A 27-bp homopurine-homopyrimidine mirror repeat adopts a triple-helical H-DNA conformation under mildly acidic conditions. This H-DNA seems to account for the S1 hypersensitivity of the promoter in vitro, since the observed pattern of S1 hypersensitivity at a single base level fits well with the H-DNA formation. Under conditions of neutral pH we have detected Z-DNA created by a (CG)5-stretch, located adjacent to the homopurine-homopyrimidine mirror repeat. The ability of the promoter DNA segment to form non-B structures has implications for models of gene regulation.


CW 50W/M2 = 10.9 diode laser source by spectral beam combining based on a transmission grating.
An external cavity structure based on the -1st transmission grating is introduced to spectral beam combining a 970 nm diode laser bar. A CW output power of 50.8 W, an electro-optical conversion efficiency of 45%, a spectral beam combining efficiency of 90.2% and a holistic M(2) value of 10.9 are achieved. This shows a way for a diode laser source with several KW power and diffraction-limited beam quality at the same time.


Studies of the DMP1 57-kDa functional domain both in vivo and in vitro.
Dmp1-null mice and patients with mutations in dentin matrix protein 1 (DMP1) resulting in autosomal recessive hypophosphatemic rickets display similar skeletal defects. As mutations were observed in the last 18 amino acids of DMP1 in 1 subset of patients and as fragments of intact DMP1, a 37-kDa N-terminal and a 57-kDa C-terminal fragment, have been purified from bone and dentin, we hypothesized that the cleaved 57-kDa C-terminal fragment is the essential functional domain of DMP1. To test this hypothesis, different forms of recombinant DMP1 were expressed in 293EBNA, CHO and 2T3 cells. The results showed that DMP1 was processed into a 37-kDa N-terminal and a 57-kDa C-terminal fragment in vitro in all cell lines examined. DMP1 processing in CHO cells was blocked by a furin protease inhibitor, decanoyl-Arg-Val-Lys-Arg-chloromethyl ketone, in a dose-dependent manner. Coexpression of PHEX, a potential upstream protease, had no apparent effect on DMP1 cleavage in 293EBNA cells, suggesting that PHEX may not be required for DMP1 processing. To test the in vivo role of the C-terminal fragment, transgenic mice overexpressing full-length DMP1 or the 57-kDa fragment controlled by the 3.6-kb Col1 promoter were generated. Overexpression of these transgenes had no effect on the wild-type skeleton, but on the Dmp1-null background showed expression in the osteoblast layer and throughout the bone matrix leading to the rescue of the null bone phenotype. This suggests that the 57-kDa C-terminal fragment may be able to recapitulate the function of intact DMP1 in vivo.


A multi-scale approach reveals that NF-κB cRel enforces a B-cell decision to divide.
Understanding the functions of multi-cellular organs in terms of the molecular networks within each cell is an important step in the quest to predict phenotype from genotype. B-lymphocyte population dynamics, which are predictive of immune response and vaccine effectiveness, are determined by individual cells undergoing division or death seemingly stochastically. Based on tracking single-cell time-lapse trajectories of hundreds of B cells, single-cell transcriptome, and immunofluorescence analyses, we constructed an agent-based multi-modular computational model to simulate lymphocyte population dynamics in terms of the molecular networks that control NF-κB signaling, the cell cycle, and apoptosis. Combining modeling and experimentation, we found that NF-κB cRel enforces the execution of a cellular decision between mutually exclusive fates by promoting survival in growing cells. But as cRel deficiency causes growing B cells to die at similar rates to non-growing cells, our analysis reveals that the phenomenological decision model of wild-type cells is rooted in a biased race of cell fates. We show that a multi-scale modeling approach allows for the prediction of dynamic organ-level physiology in terms of intra-cellular molecular networks.


Presence of Concurrent Derangements of Liver Function Tests in Type 2 Diabetes and Their Relationship with Glycemic Status: A Retrospective Observational Study from Meghalaya.
The liver plays a pivotal role in carbohydrate metabolism. Therefore, functional state of the liver in patients with diabetes is of interest. The objectives of the current study were to (i) identify co-existent biochemical derangements of liver function tests (LFTs) in type 2 diabetes and (ii) determine the association between liver function parameters and glycemic status in type 2 diabetics from Shillong, Meghalaya. Data from 320 type 2 diabetes patients were screened retrospectively for abnormalities in LFTs. Relationship of fasting serum glucose was assessed with the following tests in the LFT panel: Total bilirubin, aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), and albumin. Correlation coefficient was computed between individual LFT and fasting glucose status. These bivariate analyses were supplemented by multivariate linear regression analyses. 71.25% subjects had an abnormality in at least one LFT. Elevated ALT (46.8%) and elevated ALP (48.5%) were the most common abnormality in males and females, respectively. ALP correlated positively with fasting glucose in both sexes. AST, ALT, and ALP were found to be independent determinants of glycemic status. Derangements in liver function are widely co-existent in type 2 diabetics from Shillong. Deranged liver enzymes are associated with glycemic status. Screening for liver dysfunction in diabetics and subsequent workup may lead to the identification of hepatic co-morbidities and better management.


Physical function and fear of falling 2 years after the health-promoting randomized controlled trial: elderly persons in the risk zone.
To investigate the effects of 2 different health-promoting interventions on physical performance, fear of falling, and physical activity at 3-month, 1-year, and 2-year follow-ups of the study Elderly Persons in the Risk Zone. A randomized, three-armed, single-blind, and controlled study in which 459 independent and community-dwelling people aged 80 years or older were included. A single preventive home visit including health-promoting information and advice and 4 weekly senior group meetings focused on health strategies and peer learning, with a follow-up home visit, were compared with control. Functional balance, walking speed, fear of falling, falls efficacy, and frequency of physical activities were measured 3 months, 1 year, and 2 years after baseline. There were no or limited differences between the groups at the 3-month and 1-year follow-ups. At 2 years, the odds ratio for having a total score of 48 or more on the Berg Balance scale compared with control was 1.80 (confidence interval 1.11-2.90) for a preventive home visit and 1.96 (confidence interval 1.21-3.17) for the senior meetings. A significantly larger proportion of intervention participants than controls maintained walking speed and reported higher falls efficacy. At 1 and 2 years, a significantly higher proportion of intervention participants performed regular physical activities than control. Both a preventive home visit and senior meetings reduced the deterioration in functional balance, walking speed, and falls efficacy after 2 years. The long-term effects of both interventions indicate a positive impact on postponement of physical frailty among independent older people.


Arthroscopic surgery of the ankle.
Ankle arthroscopy is a useful technique, especially for intra-articular problems and osteocartilagenous lesions. Indications for this diagnostic and therapeutic technique include: ankle sprain unresponsive to the usual course of treatment, complaints of intermittent locking, pain and clicking, limitation of motion, and painful conditions with no obvious etiology (especially in a worker's compensation beneficiary). A review of arthroscopic, radiographic, and clinical data of all patients undergoing ankle arthroscopy at our center provided the following diagnoses: talar dome osteochondral fractures, loose bodies, accessory ossicles, talar dome cyst with loose bodies, and chronic synovitis. The anatomy of ankle arthroscopy is demanding; while the technique incorporates the same instrumentation as is used for knee arthroscopy, awareness of the neurovascular structures is essential to avoid complications. Although infrequent, complications can include sinus tract formation, sensory nerve damage, synovitis, infection, instrument breakage, and calf compartment syndromes due to extravasation of irrigation fluid.


Epigenetic research in multiple sclerosis: progress, challenges, and opportunities.
Multiple sclerosis (MS) is a chronic inflammatory and demyelinating disease of the central nervous system. MS likely results from a complex interplay between predisposing causal gene variants (the strongest influence coming from HLA class II locus) and environmental risk factors such as smoking, infectious mononucleosis, and lack of sun exposure/vitamin D. However, little is known about the mechanisms underlying MS development and progression. Moreover, the clinical heterogeneity and variable response to treatment represent additional challenges to a comprehensive understanding and efficient treatment of disease. Epigenetic processes, such as DNA methylation and histone posttranslational modifications, integrate influences from the genes and the environment to regulate gene expression accordingly. Studying epigenetic modifications, which are stable and reversible, may provide an alternative approach to better understand and manage disease. We here aim to review findings from epigenetic studies in MS and further discuss the challenges and clinical opportunities arising from epigenetic research, many of which apply to other diseases with similar complex etiology. A growing body of evidence supports a role of epigenetic processes in the mechanisms underlying immune pathogenesis and nervous system dysfunction in MS. However, disparities between studies shed light on the need to consider possible confounders and methodological limitations for a better interpretation of the data. Nevertheless, translational use of epigenetics might offer new opportunities in epigenetic-based diagnostics and therapeutic tools for a personalized care of MS patients.


CT and MR imaging of massive macronodular adrenocortical disease: a rare cause of autonomous primary adrenal hypercortisolism.
We studied four patients with adrenocorticotropic hormone (ACTH)-independent hypercortisolism due to bilateral massive enlargement of the adrenal glands. The combined weight of the adrenal glands ranged from 69 to 149 g and the adrenal cortex was replaced in three of four patients by multiple nodules ranging from microscopic to 4 cm in diameter. One patient had massive diffuse enlargement. All patients had low or undetectable levels of serum ACTH, absence of petrosal sinus to peripheral gradients of ACTH in bilateral samples from the inferior petrosal sinuses before and after stimulation by corticotropin releasing hormone, and absence of an adenoma on MR imaging of the pituitary gland. The marked degree of adrenocortical enlargement and absence of ACTH dependency separates this massive macronodular disease from the more common ACTH-dependent macronodular hyperplasia encountered in older patients with pituitary-dependent Cushing disease. All patients required bilateral adrenalectomy to control hypercortisolism. We present the spectrum of nodular adrenal disease associated with hypercortisolism and a differential diagnosis based on morphologic criteria.


Antiseizure Effects of Peganum harmala L. and Lavandula angustifolia.
Peganum harmala L. and Lavandula angustifolia are two traditional herbs with probable antiseizure effects. This study evaluated the effects of these two herbal extracts on pentylenetetrazol- (PTZ-) induced seizures in mice. We prepared hydroalcoholic extracts using P. harmala seeds and the aerial parts of L. angustifolia and then randomly divided 190 mice into 19 groups. Normal saline (10 mg/kg), diazepam (2 mg/kg), P. harmala (2.5, 5, 10, 15, 30, 45, and 60 mg/kg), and L. angustifolia (200, 400, 600, and 800 mg/kg) were intraperitoneally (IP) administrated 30 min before an IP administration of PTZ (90 mg/kg). Animals were observed for behavioral changes for one hour. In addition, the effects of flumazenil and naloxone on the antiseizure activity of P. harmala and L. angustifolia were assessed. P. harmala showed antiseizure activity at the dose of 10 mg/kg; it prolonged the seizure latency and decreased the seizure duration. The mortality protection rate was 90% for this herbal extract. L. angustifolia (600 mg/kg) prolonged the seizure latency and decreased both seizure duration and mortality. Neither flumazenil nor naloxone significantly reversed the antiseizure activities of P. harmala and L. angustifolia. In mice, the hydroalcoholic extracts of P. harmala and L. angustifolia showed antiseizure activity against PTZ-induced seizures. We could not delineate the exact antiseizure mechanisms of these extracts in the current study.


Pathological complete response after preoperative systemic therapy and outcome: relevance of clinical and biologic baseline features.
In order to evaluate the outcome of patients with breast cancer according to response after primary therapy and according to clinical and biologic baseline features, we identified patients who were treated with preoperative therapy and who underwent surgery at the European Institute of Oncology (IEO), Milan, Italy, between 1995 and 2006. The outcome of patients who achieved pathological complete remission (pCR) and patients with residual disease (RD) at final surgery was analyzed. Of the 687 patients treated with preoperative therapy, we identified 82 patients who achieved pCR (12%) and 605 patients with RD (88%). A statistically significant difference in disease-free survival (DFS), distant disease-free survival (DDFS), and overall survival (OS) was observed for patients with pCR compared with those who had RD (5 year DFS 73% vs. 59% P = 0.029; 5 year DDFS 81% vs. 72% P = 0.085; 5 year OS 88% vs. 77% P = 0.033). At the multivariate analysis, for patients achieving pCR, large tumor size (> 5 cm) correlated with worse DFS (HR 3.18; 95% CI 1.34-7.51); clinical nodal involvement was associated with poorer DFS and DDFS (HR 6.94; 95% CI 1.62-29.73 and HR 9.87 95% CI 1.29-75.53, respectively). pCR after preoperative systemic therapy correlated with significant improved outcome. A substantial rate of relapse was observed for patients with large tumors and with clinical nodal involvement at baseline. Further improvement in adjuvant treatment might be warranted.


The ecology and epidemiology of malaria parasitism in wild chimpanzee reservoirs.
Chimpanzees (Pan troglodytes) harbor rich assemblages of malaria parasites, including three species closely related to P. falciparum (sub-genus Laverania), the most malignant human malaria parasite. Here, we characterize the ecology and epidemiology of malaria infection in wild chimpanzee reservoirs. We used molecular assays to screen chimpanzee fecal samples, collected longitudinally and cross-sectionally from wild populations, for malaria parasite mitochondrial DNA. We found that chimpanzee malaria parasitism has an early age of onset and varies seasonally in prevalence. A subset of samples revealed Hepatocystis mitochondrial DNA, with phylogenetic analyses suggesting that Hepatocystis appears to cross species barriers more easily than Laverania. Longitudinal and cross-sectional sampling independently support the hypothesis that mean ambient temperature drives spatiotemporal variation in chimpanzee Laverania infection. Infection probability peaked at ~24.5 °C, consistent with the empirical transmission optimum of P. falciparum in humans. Forest cover was also positively correlated with spatial variation in Laverania prevalence, consistent with the observation that forest-dwelling Anophelines are the primary vectors. Extrapolating these relationships across equatorial Africa, we map spatiotemporal variation in the suitability of chimpanzee habitat for Laverania transmission, offering a hypothetical baseline indicator of human exposure risk.


Rational and design of an individual participant data meta-analysis of spinal manipulative therapy for chronic low back pain-a protocol.
Chronic low back pain (LBP) is the leading cause of pain and disability, resulting in a major socioeconomic impact. The Cochrane Review which examined the effect of spinal manipulative therapy (SMT) for chronic LBP concluded that SMT is moderately effective, but was based on conventional meta-analysis of aggregate data. The use of individual participant data (IPD) from trials allows for a more precise estimate of the treatment effect and has the potential to identify moderators and/or mediators. The aim is (1) to assess the overall treatment effect of SMT for primary and secondary outcomes in adults with chronic LBP, (2) to determine possible moderation of baseline characteristics on treatment effect, (3) to identify characteristics of intervention (e.g., manipulation/mobilization) that influence the treatment effect, and (4) to identify mediators of treatment effects. All trials included in the Cochrane Review on SMT for chronic LBP will be included which were published after the year 2000, and the search will be updated. No restrictions will be placed on the type of comparison or size of the study. Primary outcomes are pain intensity and physical functioning. A dataset will be compiled consisting of individual trials and variables included according to a predefined coding scheme. Variables to be included are descriptive of characteristics of the study, treatment, comparison, participant characteristics, and outcomes at all follow-up periods. A one-stage approach with a mixed model technique based on the intention-to-treat principle will be used for the analysis. Subsequent analyses will focus on treatment effect moderators and mediators. We will analyze IPD for LBP trials in which SMT is one of the interventions. IPD meta-analysis has been shown to be more reliable and valid than aggregate data meta-analysis, although this difference might also be attributed to the number of studies that can be used or the amount of data that can be utilized. Therefore, this project may identify important gaps in our knowledge with respect to prognostic factors of treatment effects. PROSPERO CRD42015025714.


Free radical induced increase in protein carbonyl is attenuated by low dose of adenosine in hippocampus and mid brain: implication in neurodegenerative disorders.
There is strong evidence that oxidative stress participates in the etiology of neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease. In the previous studies we have already shown that a combination of alpha-tocopherol and ascorbic acid protect neurons against tert-butyl hydroperoxide (t-BuOOH) induced neurotoxicity in different brain regions including hippocampus and mid brain. In this work, we examined the neuroprotective effect of low dose of adenosine against protein oxidation (protein carbonyls) in parallel with the level of reduced glutathione (GSH) in hippocampus and mid brain regions of mouse brain. The t-BuOOH was injected intraperitoneally in three concentrations (50, 100, 150 mg/kg b.w.) for 10 days. Results showed dose dependent increase in protein carbonyl (PC) in hippocampus and mid brain region. This increase was accompanied by a significant (p < 0.05) decline in GSH content in both brain regions of t-BuOOH treated mice. Adenosine (1 mg/kg b.w.) protected both hippocampus and mid brain neurons against protein oxidation as evidenced by reduction in protein carbonyl content. The GSH content was significantly (p < 0.05) increased after the treatment of adenosine in both brain regions. These data show that prior treatment with low dose of adenosine attenuates the oxidative protein damage with parallel increase in the GSH level in hippocampus and mid brain of t-BuOOH induced mice.


Cytogenetics and molecular genetics of myelodysplastic neoplasms.
According to the 2022 World Health Organization (WHO) Classification (5th edition), the term myelodysplastic neoplasms (abbreviated MDS) has been introduced to replace myelodysplastic syndromes. MDS are a group of clonal hematopoietic stem cell diseases characterized by cytopenia(s), dysplasia in one or more of lineages, ineffective hematopoiesis, and an increased risk of progression to bone marrow failure or to acute myeloid leukemia (AML). Current NCCN guidelines and recent review articles have provided in depth discussion on the clinical diagnosis and management of MDS. This review will focus on discussion of the WHO and International Consensus Classification (ICC) updates on the role of cytogenetics and molecular genetics in the diagnosis and risk stratification of MDS.


Complex and dynamic transcriptional changes allow the helminth Fasciola gigantica to adjust to its intermediate snail and definitive mammalian hosts.
The tropical liver fluke, Fasciola gigantica causes fasciolosis, an important disease of humans and livestock. We characterized dynamic transcriptional changes associated with the development of the parasite in its two hosts, the snail intermediate host and the mammalian definitive host. Differential gene transcription analysis revealed 7445 unigenes transcribed by all F. gigantica lifecycle stages, while the majority (n = 50,977) exhibited stage-specific expression. Miracidia that hatch from eggs are highly transcriptionally active, expressing a myriad of genes involved in pheromone activity and metallopeptidase activity, consistent with snail host finding and invasion. Clonal expansion of rediae within the snail correlates with increased expression of genes associated with transcription, translation and repair. All intra-snail stages (miracidia, rediae and cercariae) require abundant cathepsin L peptidases for migration and feeding and, as indicated by their annotation, express genes putatively involved in the manipulation of snail innate immune responses. Cercariae emerge from the snail, settle on vegetation and become encysted metacercariae that are infectious to mammals; these remain metabolically active, transcribing genes involved in regulation of metabolism, synthesis of nucleotides, pH and endopeptidase activity to assure their longevity and survival on pasture. Dramatic growth and development following infection of the mammalian host are associated with high gene transcription of cell motility pathways, and transport and catabolism pathways. The intra-mammalian stages temporally regulate key families of genes including the cathepsin L and B proteases and their trans-activating peptidases, the legumains, during intense feeding and migration through the intestine, liver and bile ducts. While 70% of the F. gigantica transcripts share homology with genes expressed by the temperate liver fluke Fasciola hepatica, gene expression profiles of the most abundantly expressed transcripts within the comparable lifecycle stages implies significant species-specific gene regulation. Transcriptional profiling of the F. gigantica lifecycle identified key metabolic, growth and developmental processes the parasite undergoes as it encounters vastly different environments within two very different hosts. Comparative analysis with F. hepatica provides insight into the similarities and differences of these parasites that diverged > 20 million years ago, crucial for the future development of novel control strategies against both species.


Facilitation of quadriceps activation following a concentrically controlled knee flexion movement: the influence of transition rate.
Single group repeated measures design. To determine if the rate of transition between knee flexion and extension influences the subsequent concentric activation of the quadriceps and knee extension torque during reciprocal movements. Preloading a muscle by stretching, a prior isometric or eccentric muscle action, or a prior movement controlled concentrically by the antagonist muscle group increases the maximal torque-generating capability of the agonist. We hypothesized that the rate of transition from the prior movement may be the critical factor that influences the degree of muscle facilitation and torque potentiation. Rapid reversal of antagonistic movements has been postulated as a potential facilitatory mechanism. Knee extension torque and electromyographic (EMG) amplitude (dependent variables) from 2 of the vasti muscles were recorded while subjects (N = 20; 12 men, 8 women, mean age, 28.5+/-2.68 years) maximally activated their quadriceps at 3 constant angular velocities, 100 degrees/s, 200 degrees/s, and 300 degrees/s, and 2 preload conditions, SLOW and RAPID (independent variables). In the SLOW transition condition, subjects actively flexed their knee to 110 degrees from an extended position, paused in this position for 3 seconds, and then extended to 0 degrees. In the RAPID transition condition, the same movement from knee flexion to extension was performed without a pause. Peak torque, the root-mean-square (RMS) average, peak (peak rectified and smoothed), and initial (100 milliseconds prior to torque onset) EMG amplitudes were all significantly greater during the RAPID transition condition. Peak torque decreased with increasing movement velocity. There were no interactions between the preload conditions and angular velocity on peak torque or the EMG amplitude variables. There was also no influence of velocity on the EMG amplitude variables. The effect of preloading the quadriceps by prior concentric activation of the hamstrings is dependent on the rate of transition between the flexion and extension movements and is due primarily to neural facilitation.


The effect of resveratrol on lifespan depends on both gender and dietary nutrient composition in Drosophila melanogaster.
Resveratrol, a polyphenolic compound, has been shown to extend lifespan in different organisms. Emerging evidence suggests that the prolongevity effect of resveratrol depends on dietary composition. However, the mechanisms underlying the interaction of resveratrol and dietary nutrients in modulating lifespan remain elusive. Here, we investigated the effect of resveratrol on lifespan of Drosophila melanogaster fed diets differing in the concentrations of sugar, yeast extract, and palmitic acid representing carbohydrate, protein, and fat, respectively. Resveratrol at up to 200 μM in diets did not affect lifespan of wild-type female flies fed a standard, restricted or high sugar-low protein diet, but extended lifespan of females fed a low sugar-high protein diet. Resveratrol at 400 μM extended lifespan of females fed a high-fat diet. Lifespan extension by resveratrol was associated with downregulation of genes in aging-related pathways, including antioxidant peroxiredoxins, insulin-like peptides involved in insulin-like signaling and several downstream genes in Jun-kinase signaling involved in oxidative stress response. Furthermore, resveratrol increased lifespan of superoxide dismutase 1 (sod1) knockdown mutant females fed a standard or high-fat diet. No lifespan extension by resveratrol was observed in wild-type and sod1 knockdown males under the culture conditions in this study. Our results suggest that the gender-specific prolongevity effect of resveratrol is influenced by dietary composition and resveratrol promotes the survival of flies by modulating genetic pathways that can reduce cellular damage. This study reveals the context-dependent effect of resveratrol on lifespan and suggests the importance of dietary nutrients in implementation of effective aging interventions using dietary supplements.


Medical care costs for diabetes associated with health disparities among adult Medicaid enrollees in North Carolina.
Health disparities for many diseases are large and long-standing in North Carolina and the nation. This study examines medical care costs for diabetes associated with health disparities among adults (age, > or =78 years) enrolled in Medicaid in North Carolina during state fiscal year (SFY) 2007-2008 (i.e., July 7, 2007, through June 30, 2008). North Carolina Medicaid paid claims and enrollment data were used to calculate the prevalence of and medical care expenditures for diabetes among adult Medicaid enrollees overall and by white, African American, and American Indian race. The impacts of racial and economic health disparities on medical care costs for diabetes were determined by first calculating the proportionate differences between the diabetes prevalence for whites, African Americans, and American Indians enrolled in Medicaid and the diabetes prevalence among all whites in North Carolina. Then it was assumed that medical care costs for white, African American, and American Indian Medicaid recipients could be reduced by the same proportion if the overall prevalence among whites was achieved. The diabetes prevalence among adult Medicaid enrollees was 75.7%, compared with 9.1% for all North Carolina adults. During SFY 2007-2008, the state Medicaid program in North Carolina spent $525 million for diabetes-related medical care and prescription drugs among adults. An estimated $225 million in diabetes-related expenditures could be saved each year by the North Carolina Medicaid program if both racial and economic disparities in the diabetes prevalence were eliminated. We did not have data on non-Medicaid paid health care expenditures for the Medicaid enrollees in our study. The costs of interventions to eliminate health disparities associated with diabetes are not included in the calculation of the potential savings. The diabetes prevalence in the Medicaid population is much greater than that for all North Carolinians, and the Medicaid costs associated with this elevated prevalence are large. North Carolina health-policy makers and health-program managers should carefully evaluate investments in interventions to reduce these race- and economic-based differences in diabetes prevalence, which could potentially reduce Medicaid costs.


Postoperative Clinical Course and Opioid Consumption Following Repair of Congenital Diaphragmatic Hernia: Open Versus Thoracoscopic Techniques.
Introduction: Minimally invasive surgical (MIS) approaches for thoracic procedures in adults result in an improved postoperative course with less pain, but there are limited data on similar procedures in neonates. We aimed to evaluate postoperative opioid consumption and pain management practices in neonates and infants following MIS versus open repair of congenital diaphragmatic hernia (CDH). Materials and Methods: This was an IRB approved, retrospective study from 2012 to 2016. Demographic data, intraoperative analgesic regimen, total 7-day postoperative opioid consumption, and use of adjunctive pain medications were compared by surgery type (open versus MIS). Secondary measures included time to tracheal extubation, oral feeds, and discharge home. Results: The study cohort included 28 patients (13 female, median age 5 days, average gestational age 39 weeks, and weight 3 kg). MIS was performed in 8 patients. In the first 7 postoperative days, the median postoperative opioid consumption was 0.3 mg/kg of oral morphine equivalents (interquartile range [IQR] 0.2, 18.3) in the MIS group versus 32.3 mg/kg (IQR 9.9, 53.6) in the open group (95% CI of differences in medians: 8.2-42.9; P = .006). No difference was noted in intraoperative opioid administration. Among secondary outcomes, length of stay was significantly longer in the open group. Conclusions: Although several factors may impact the hospital course of neonates with CDH, we found that patients had a more than 100-fold difference in median opioid consumption following repair with MIS versus an open approach. The study also noted significant variation in analgesic regimens suggesting other avenues for improved care of postsurgical neonates.


Noradrenergic alpha 1 and alpha 2 receptors: light microscopic autoradiographic localization.
[3H]WB-4101 and p-[3H]aminoclonidine were used for light microscopic autoradiographic localization of alpha 1 and alpha 2 adrenergic receptors, respectively, in the rat brain. The binding of these ligands to slide-mounted tissue sections had all of the characteristics associated with alpha 1 and alpha 2 receptors. It was saturable with appropriate kinetic constants and was blocked only by other alpha-adrenergic drugs with the appropriate potency. Autoradiographic studies revealed a distribution of alpha-adrenergic receptors throughout the nervous system. Certain areas had elevated levels. These included parts of the olfactory bulb and nucleus, parts of the cerebral cortex and dentate gyrus, the more medial portions of the hypothalamus and thalamus, the locus ceruleus and nucleus tractus solitarii, and parts of the spinal cord. In certain areas, the distribution of alpha 1 and alpha 2 receptors was markedly different. These results provide some rational basis for the observed actions of alpha-adrenergic drugs on the central nervous system. For example, the finding of high densities of alpha 2 receptors in the nucleus tractus solitarii is most likely related to its antihypertensive action. The observed codistribution of alpha 2 receptors with opiate receptors would provide an explanation of the observation that alpha 2 agonists block opiate withdrawal. The results are also discussed in relationship to the anatomy of catecholamine systems in the brain.


Predictors of post-COVID-19 and the impact of persistent symptoms in non-hospitalized patients 12 months after COVID-19, with a focus on work ability.
Better knowledge of long-term symptoms following coronavirus disease 2019 (COVID-19), the so-called post-COVID-19, in non-hospitalized patients is needed. The aim of this study was to study persisent symptoms up to 12 months after COVID-19 in non-hospitalized patients and their impact on work ability. We also investigated predictors of persistent symptoms. This study encompassed non-hospitalized adult subjects with a COVID-19 infection confirmed via positive nasopharyngeal swab polymerase chain reaction test during the first wave of the pandemic in Uppsala, Sweden. In total, 566 subjects were sent a survey via e-mail or post with an invitation to participate in the survey 12 months post-diagnosis. The majority of subjects were healthcare workers, as this group was prioritized for testing. A total of 366 subjects responded, with 47% reporting persistent symptoms 12 months after their COVID-19 diagnosis. The most commonly reported symptoms at this time were impaired sense of smell and/or taste and fatigue. Among the predictors of persistent symptoms were being born abroad, lower physical fitness compared with peers before COVID-19, body mass index >25 kg/m2, cooccurrence of hypertension and chronic pain, and having more than seven of the general COVID-19 symptoms at the onset. Respondents with symptoms after 12 months self-reported negatively about their general health and work ability. This study indicated that many people who had mild COVID-19 might have a variety of long-term symptoms. It highlights the importance of considering work ability after mild COVID-19.


Relative efficiency of warbled-tone and conventional pure-tone testing with children.
Thirty-four normal-hearing 4-year-old children were tested with conventional steady-state pure tones and with warbled tones to compare efficiency of the stimuli. Three measures of efficiency were used: (1) amount of time to train a child to perform the experimental task; (2) amount of time required to obtain thresholds; and (3) number of ascents used for obtaining thresholds. The strong implication by the literature that warbled tones would result in greater clinical efficiency was not supported by the comparisons of the experimental measures. It was concluded that convincing evidence to persuade the audiologist to select warbled over conventional steady-state pure tones for testing children was lacking.


[Nature of centrogenic reactions developing during surgical interventions on the brain].
The nature of hemodynamic reactions and EEG changes developing during surgery is discussed, based on analysis of 392 interventions for removal of brain tumors. Two main types of centrogenic reactions are distinguished: (1) connected with the direct effect on brain centers engaged in cardiovascular regulation and (2) caused by stimulation of the median structures of the brain, most often caused by hyperperfusion in different compartments of the brain. Use of anesthesiological method limiting the functional relations between the brain stem and forebrain notably decreases the incidence of centrogenic reactions during neurosurgical interventions.


Basal ganglia physiology and deep brain stimulation.
Despite improvements in anatomic imaging of the basal ganglia, microelectrode recording is still an invaluable tool in locating appropriate targets for neurosurgical intervention. These recording also provide an unparalleled opportunity to study the pathophysiological aspects of diseases. This article reviews the principles of microelectrode recording in functional neurosurgery and discusses the pathologic neurophysiologic findings commonly encountered. It also highlights some of the potential mechanisms of action of both dopaminergic drugs and deep brain stimulation. In addition we review the recent work on pedunculopontine nucleus neurophysiology and trials of deep brain stimulation in that region for gait disturbances in Parkinson's disease.


The Stress-Chip: A microfluidic platform for stress analysis in Caenorhabditis elegans.
An organism's ability to mount a physiological response to external stressors is fundamental to its interaction with the environment. Experimental exploration of these interactions benefits greatly from the ability to maintain tight control of the environment, even under conditions in which it would be normal for the subject to flee the stressor. Here we present a nematode research platform that pairs automated image acquisition and analysis with a custom microfluidic device. This platform enables tight environmental control in low-density, single-worm arenas, which preclude animal escape while still allowing a broad range of behavioral activities. The platform is easily scalable, with two 50 arena arrays per chip and an imaging capacity of 600 animals per scanning device. Validating the device using dietary, osmotic, and oxidative stress indicates that it should be of broad use as a research platform, including eventual adaptation for additional stressors, anthelmintic-drug screening, and toxicology studies.


Effectiveness of Group Problem Management Plus (Group-PM+) for adults affected by humanitarian crises in Nepal: study protocol for a cluster randomized controlled trial.
Globally, the lack of availability of psychological services for people exposed to adversities has led to the development of a range of scalable psychological interventions with features that enable better scale-up. Problem Management Plus (PM+) is a brief intervention of five sessions that can be delivered by non-specialists. It is designed for people in communities in low- and middle-income countries (LMIC) affected by any kind of adversity. Two recent randomized controlled trials in Pakistan and Kenya demonstrated the effectiveness of individually delivered PM+. A group version of PM+ has been developed to make the intervention more scalable and acceptable. This paper describes the protocol for a cluster randomized controlled trial (c-RCT) on locally adapted Group PM+ in Nepal. This c-RCT will compare Group PM+ to enhanced usual care (EUC) in participants with high levels of psychological distress recruited from the community. The study is designed as a two-arm, single-blind c-RCT that will be conducted in a community-based setting in Morang, a flood affected district in Eastern Nepal. Randomization will occur at ward level, the smallest administrative level in Nepal, with 72 enrolled wards allocated to Group PM+ or to EUC (ratio 1:1). Group PM+ consists of five approximately 2.5-h sessions, in which participants are taught techniques to manage their stressors and problems, and is delivered by trained and supervised community psychosocial workers (CPSWs). EUC consists of a family meeting with (a) basic information on adversity and mental health, (b) benefits of getting support, (c) information on seeking services from local health facilities with mhGAP-trained staff. The primary outcome measure is levels of individual psychological distress at endline (equivalent to 20 ± 1 weeks after baseline), measured by the General Health Questionnaire (GHQ-12). Secondary outcome measures include levels of functioning, depressive symptoms, post-traumatic stress disorder symptoms, levels of social support, somatic symptoms, and ways of coping. We hypothesize that skills acquired will mediate any impact of the intervention. This c-RCT will contribute to the growing evidence-base for transdiagnostic psychological interventions delivered by non-specialists for people in communities affected by adversity. If Group PM+ is proven effective, the intervention manual will be released for use, giving the opportunity for further adaptation and implementation of the intervention in diverse settings with communities that require better access to psychological interventions. ClinicalTrials.gov, NCT03747055.


Immunohistochemistry of gastric carcinomas and associated diseases: novel distribution of carcinoembryonic antigen and secretory component on the surface of gastric cancer cells.
Carcinoembryonic antigen (CEA) and secretory component (SC) were localized by peroxidase-labeled antibody immunocytochemistry in normal and abnormal human gastric mucosa. In normal epithelium, both glycoproteins were absent or only faintly present, but in intestinal metaplasia and carcinoma both were prominently present. CEA and SC on the surfaces of metaplastic epithelial cells were polarized. That is, CEA was expressed only on the microvillous surface and SC was expressed only on the basolateral surface. In gastric cancer, CEA and SC were distributed over the entire surface of the neoplastic cells. Thus, deviations from the normal differentiation and maturation of gastric epithelial cells were accompanied by abnormalities in surface expression of CEA and SC. These observations, together with compatible observations previously made in colonic neoplasia (DJ Ahnen, PK Nakane, and WR Brown, Cancer 49:2077, 1982), suggest that loss of polarity of surface membrane components is a characteristic of neoplastic epithelial cells.


Therapy for septic arthritis.
The usefulness of the results of serial cultures of synovial fluid in assessing the response to medical therapy of acute bacterial joint infections was examined. A five-year retrospective survey disclosed 32 synovial fluid culture-proved cases of septic arthritis in adults. Thirteen of the patients had well-documented evidence of (1) duration of articular symptoms (presumably reflecting the presence of infection) and (2) subsequent time required to sterilize the synovial fluids (as determined by serial synovial fluid cultures). We found a positive correlation between duration of symptoms and length of time during which serial cultures of synovial fluids remained positive after initiation of treatment. In the seven patients with good outcome, cultures remained positive for 2.4+/-1.9 days, as opposed to 8.2+/-2.4 days in the six patients with poor results. The time required to sterilize the synovial fluid appears to be a reliable indicator of response to therapy and an accurate predictor of outcome. Our data show that the rapidity in achieving synovial fluid sterility is of paramount importance in the successful treatment of infection, preservation of articular integrity, and restoration of joint function.


The serine-threonine kinase PIM3 is an aldosterone-regulated protein in the distal nephron.
The mineralocorticoid hormone aldosterone plays a crucial role in the control of Na+ and K+ balance, blood volume, and arterial blood pressure, by acting in the aldosterone-sensitive distal nephron (ASDN) and stimulating a complex transcriptional, translational, and cellular program. Because the complexity of the aldosterone response is still not fully appreciated, we aimed at identifying new elements in this pathway. Here, we demonstrate that the expression of the proto-oncogene PIM3 (Proviral Integration Site of Moloney Murine Leukemia Virus 3), a serine/threonine kinase belonging to the calcium/calmodulin-regulated group of kinases, is stimulated by aldosterone in vitro (mCCDcl1 cells), ex vivo (mouse kidney slices), and in vivo in mice. Characterizing a germline Pim3-/- mouse model, we found that these mice have an upregulated Renin-Angiotensin-Aldosterone System (RAAS), with high circulating aldosterone and plasma renin activity levels on both standard or Na+ -deficient diet. Surprisingly, we did not observe any obvious salt-losing phenotype in Pim3 KO mice as shown by normal blood pressure, plasma and urinary electrolytes, as well as unchanged expression levels of the major Na+ transport proteins. These observations suggest that the potential effects of the loss of the Pim3 gene are physiologically compensated. Indeed, the 2 other family members of the PIM kinase family, PIM1 and PIM2 are upregulated in the kidney of Pim3-/- mice, and may therefore be involved in such compensation. In conclusion, our data demonstrate that the PIM3 kinase is a novel aldosterone-induced protein, but its precise role in aldosterone-dependent renal homeostasis remains to be determined.


Breast tumour imaging using incomplete circular orbit pinhole SPET: a phantom study.
Improvements in 99Tcm-sestamibi breast lesion visualization using single photon emission tomography (SPET) may help define the clinical role of this technique alongside X-ray mammography in the diagnosis and management of breast cancer. Pinhole SPET offers the advantages of high resolution and sensitivity when compared to conventional parallel-beam collimation for sources located near the pinhole aperture. In this work, the potential of incomplete (180 degrees) circular orbit (ICO) SPET with pinhole collimation is investigated as a means to visualize small (6.4 and 9.6 mm diameter) spherical simulated tumours, at clinical count densities and tumour-to-background ratios, in a breast phantom. ICO pinhole SPET is compared to complete circular orbit (CCO) pinhole SPET for reference, and planar breast imaging (scintimammography) using parallel-beam and pinhole collimators. A prototype box-shaped pinhole collimator with a 4 mm diameter circular aperture was used to acquire projections of an 890 ml breast phantom both in isolation and mounted on a cylinder filled with a mixture of 99Tcm-pertechnetate and water. A heart phantom containing 99Tcm activity in the myocardium was placed in the cylinder. Simulated tumours containing 99Tcm were placed in the breast phantom and scanned at clinically relevant count densities and scan times with tumour-to-normal tissue concentration ratios of 5.0:1 (9.6 mm sphere) and 7.7:1 (6.4 mm sphere). Phantom data were reconstructed using pinhole filtered backprojection (FBP) and maximum likelihood-expectation maximization (ML-EM). The tumours were not visualized with scintimammography, in which lesion contrast and signal-to-noise were estimated from region of interest analysis to be < 2% and 0.01, respectively. Average (over lesion size and scan time) contrast and signal-to-noise in the ICO (CCO) SPET images were 33% and 1.72 (34% and 1.3), respectively. These values indicate that ICO pinhole SPET has the potential to improve visualization of small (< 10 mm) breast tumours when compared with scintimammography, which may be beneficial for the early classification of cancers of the breast.


Continuous activation of the CD122/STAT-5 signaling pathway during selection of antigen-specific regulatory T cells in the murine thymus.
Signaling events affecting thymic selection of un-manipulated polyclonal natural CD25(+)foxp3(+) regulatory T cells (nTreg) have not been established ex vivo. Here, we report a higher frequency of phosphorylated STAT-5 (pSTAT-5) in nTreg cells in the adult murine thymus and to a lesser extent in the periphery, compared to other CD4(+)CD8(-) subsets. In the neonatal thymus, the numbers of pSTAT-5(+) cells in CD25(+)foxp3(-) and nTreg cells increased in parallel, suggesting that pSTAT-5(+)CD25(+)foxp3(-) cells might represent the precursors of foxp3(+) regulatory T cells. This "specific" pSTAT-5 expression detected in nTreg cells ex vivo was likely due to a very recent signal given by IL-2/IL-15 cytokines in vivo since (i) it disappeared rapidly if cells were left unstimulated in vitro and (ii) was also observed if total thymocytes were stimulated in vitro with saturating amounts of IL-2 and/or IL-15 but not IL-7. Interestingly, STAT-5 activation upon IL-2 stimulation correlated better with foxp3 and CD122 than with CD25 expression. Finally, we show that expression of an endogenous superantigen strongly affected the early Treg cell repertoire but not the proportion of pSTAT-5(+) cells within this repertoire. Our results reveal that continuous activation of the CD122/STAT-5 signaling pathway characterize regulatory lineage differentiation in the murine thymus.


Genomic Comparison of Indigenous African and Northern European Chickens Reveals Putative Mechanisms of Stress Tolerance Related to Environmental Selection Pressure.
Global climate change is increasing the magnitude of environmental stressors, such as temperature, pathogens, and drought, that limit the survivability and sustainability of livestock production. Poultry production and its expansion is dependent upon robust animals that are able to cope with stressors in multiple environments. Understanding the genetic strategies that indigenous, noncommercial breeds have evolved to survive in their environment could help to elucidate molecular mechanisms underlying biological traits of environmental adaptation. We examined poultry from diverse breeds and climates of Africa and Northern Europe for selection signatures that have allowed them to adapt to their indigenous environments. Selection signatures were studied using a combination of population genomic methods that employed FST , integrated haplotype score (iHS), and runs of homozygosity (ROH) procedures. All the analyses indicated differences in environment as a driver of selective pressure in both groups of populations. The analyses revealed unique differences in the genomic regions under selection pressure from the environment for each population. The African chickens showed stronger selection toward stress signaling and angiogenesis, while the Northern European chickens showed more selection pressure toward processes related to energy homeostasis. The results suggest that chromosomes 2 and 27 are the most diverged between populations and the most selected upon within the African (chromosome 27) and Northern European (chromosome 2) birds. Examination of the divergent populations has provided new insight into genes under possible selection related to tolerance of a population's indigenous environment that may be baselines for examining the genomic contribution to tolerance adaptions.


Exploring Rain as Source of Biological Control Agents for Fire Blight on Apple.
Poor survival on plants can limit the efficacy of Biological Control Agents (BCAs) in the field. Yet bacteria survive in the atmosphere, despite their exposure to high solar radiation and extreme temperatures. If conditions in the atmosphere are similar to, or more extreme than, the environmental conditions on the plant surface, then precipitation may serve as a reservoir of robust BCAs. To test this hypothesis, two hundred and fifty-four rain-borne isolates were screened for in vitro inhibition of Erwinia amylovora, the causal agent of fire blight, as well as of other plant pathogenic bacteria, fungi and oomycetes. Two isolates showed strong activity against E. amylovora and other plant pathogenic bacteria, while other isolates showed activity against fungal and oomycete pathogens. Survival assays suggested that the two isolates that inhibited E. amylovora were able to survive on apple blossoms and branches similarly to E. amylovora. Pathogen population size and associated fire blight symptoms were significantly reduced when detached apple blossoms were treated with the two isolates before pathogen inoculation, however, disease reduction on attached blossoms within an orchard was inconsistent. Using whole genome sequencing, the isolates were identified as Pantoea agglomerans and P. ananatis, respectively. A UV-mutagenesis screen pointed to a phenazine antibiotic D-alanylgriseoluteic acid synthesis gene cluster as being at the base of the antimicrobial activity of the P. agglomerans isolate. Our work reveals the potential of precipitation as an under-explored source of BCAs, whole genome sequencing as an effective approach to precisely identify BCAs, and UV-mutagenesis as a technically simple screen to investigate the genetic basis of BCAs. More field trials are needed to determine the efficacy of the identified BCAs in fire blight control.