Journal articles on the topic 'Multi probe cut'

This paper introduces a near-field measurement system concept for the fast testing of linear arrays suited for mass production scenarios where a high number of nominally identical antennas needs to be measured. The proposed system can compute the radiation pattern, directivity and gain on the array plane, as well as the array complex feeding coefficients in a matter of seconds. The concept is based on a multi-probe antenna array arranged in a line which measures the near field of the antenna under test in its array plane. This linear measurement is postprocessed with state-of-the-art single-cut transformation techniques. To compensate the lack of full 3D information, a previous complete characterization of a “Gold Antenna” is performed. This antenna is nominally identical to the many ones that will be measured with the proposed system. Therefore, the data extracted from this full characterization can be used to complement the postprocessing steps of the single-cut measurements. An X-band 16-probe demonstrator of the proposed system is implemented and introduced in this paper, explaining all the details of its architecture and operation steps. Finally, some measurement results are given to compare the developed demonstrator with traditional anechoic measurements, and show the potential capabilities of the proposed concept to perform fast and reliable measurements.

Abstract Transient thermal dissipation (TTD) systems provide a simple way to measure xylem sap flow with dual or single Granier-type probe, which gives lower energy consumption and higher accuracy due to its lessor sensitivity to thermal interferences. A new system, TTD heat within 5 min (TTD5), proposed on the reduction of the heating duration. This evolution captured interest through decreased energy consumption and increased temporal resolution. Within our study, the first objective was to test and calibrate this new system with a single probe for young rubber tree – Hevea brasiliensis. The second objective was to explore the sources of variability in calibration such as species, individual cut-stems and probe–wood contact. The complementary species consisted of two diffuse-porous species (mango tree – Mangifera indica, eucalyptus tree – Eucalyptus camaldulensis) and one ring-porous species (teak tree – Tectonia grandis). Twenty-eight response curves were assessed over a large range of flux densities from 0.5 to 10 l dm−2 h−1. The incremental rise of temperature from 30 to 300 s (T300–30) after commencement of heating was sensitive to flux density over the complete range. Compared with the full signal at 300 s, the incremental signal markedly reduced the variability between response curves within species and between species. Moreover, a new index K2, defined as (T0 – Tu)/T0, normalized the responses between 0 and 1. However, the responses had a non-linear trend above 5 l dm−2 h−1. Within diffuse-porous wood type, the species did not differ in calibration, whereas the ring-porous species was markedly different. A sigmoid function provided the best fit for the diffuse-porous species. Individual stems were identified as the main source of within-species variability in calibration. The normalizing K2 index removed the influence of probe–wood contacts, controlled through drilling difference; however, there was still an effect of individual stems interacting with flux density (P = 0.019). Replications of cut-stems and response curves are necessary to assess a reliable averaged calibration. In conclusion, the applicability of the TTD5 system with a single probe has been confirmed and several sources of variability in calibration have been evaluated.

Purpose – This paper aims to offer the aerodynamic testing community a new procedure for manufacturing high-quality aerodynamic probes suitable for 3D flow measurements with consistent geometry and calibration by taking advantage of the additive manufacturing technology. Design/methodology/approach – The design methodology combines the advantages and flexibilities of computer aided design (CAD)/computer aided manufacturing (CAM) along with the use of computational fluid dynamics to design and analyse suitable probe shapes prior to manufacturing via rapid prototyping. Findings – A viable procedure to design and possibly batch manufacture geometrically accurate pneumatic probes with consistent calibration is shown to be possible through this work. Multi-jet modelling prototyping methods with wax-based support materials are found to be a cost-effective method when clean and long sub-millimetre pressure channels are to be cut. Originality/value – Utilisation of the geometry consistency that is made possible by 3D printing technology for the design and development of pneumatic probes is described. It is suggested that the technique could lead to batch production of identical probes, thus avoiding precious time of a skilled labourer and elaborate individual calibration requirement.

The aim of this study was to evaluate the pulpal response of dogs primary teeth to an adhesive system or to a calcium hidroxide cement after mechanic exposure of the pulp. Three mongrel dogs were used and ten class V cavities were prepared on their teeth. A mechanic pulp exposure was produced with a sterile exploratory probe in the central portion of each cavity and bleeding was controlled with dry sterile cotton pellets. Enamel, dentin and the site of the pulp exposure of five teeth were etched with 35% phosphoric acid followed by the application of an adhesive system (Scotchbond Multi-Purpose - 3M). In the other five teeth, calcium hydroxide cement (Hydro C - Dentsply) was applied on the site of the pulp exposition before application of the adhesive system (Scotchbond Multi-Purpose - 3M). All teeth were restored with a resin composite (Z-100 - 3M). After 7, 30 or 45 days the dogs were anesthetized and perfused with saline followed by a solution of neutral buffered formalin. Maxilla and mandible were sectioned into three parts and placed in a solution for demineralization. Following bone demineralization, all teeth were cut, trimmed, embedded in paraffin and longitudinally cut. Then, the teeth were stained with hematoxilin and eosin and observed under a light microscope. The results obtained with the treatments proposed in this study showed the presence and persistence of an inflammatory response of different intensities at the three experimental periods. There was no variation in the inflammatory response regarding the different treatments performed.

We study the space density evolution of active galactic nuclei (AGN) using the 610 MHz radio survey of the XXL-North field, performed with the Giant Metrewave Radio Telescope. The survey covers an area of 30.4 deg2, with a beamsize of 6.5 arcsec. The survey is divided into two parts, one covering an area of 11.9 deg2 with 1σ rms noise of 200 μJy beam−1 and the other spanning 18.5 deg2 with rms noise of 45 μJy beam−1. We extracted the catalog of radio components above 7σ. The catalog was cross-matched with a multi-wavelength catalog of the XXL-North field (covering about 80% of the radio XXL-North field) using a likelihood ratio method, which determines the counterparts based on their positions and their optical properties. The multi-component sources were matched visually with the aid of a computer code: Multi-Catalog Visual Cross-Matching. A flux density cut above 1 mJy selects AGN hosts with a high purity in terms of star formation contamination based on the available source counts. After cross-matching and elimination of observational biases arising from survey incompletenesses, the number of remaining sources was 1150. We constructed the rest-frame 1.4 GHz radio luminosity functions of these sources using the maximum volume method. This survey allows us to probe luminosities of 23 ≲ log(L1.4 GHz[W Hz−1]) ≲ 28 up to redshifts of z ≈ 2.1. Our results are consistent with the results from the literature in which AGN are comprised of two differently evolving populations, where the high luminosity end of the luminosity functions evolves more strongly than the low-luminosity end.

AbstractThe extensive ground-based spectroscopy campaign from the VIMOS Ultra-Deep Survey (VUDS), and the deep multi-wavelength photometry in three very well observed extragalactic fields (ECDFS, COSMOS, VVDS), allow us to investigate physical properties of a large sample (~4000 galaxies) of spectroscopically confirmed faint (iAB ≲ 25 mag) SFGs, with and without Lyα in emission, at z ~ 2–6. The fraction of Lyα emitters (LAEs; equivalent width (EW) ≥ 20Å) increases from ~10% at z ~ 2 to ~40% at z ~ 5–6, which is consistent with previous studies that employ higher Lyα EW cut. This increase in the LAE fraction could be, in part, due to a decrease in the dust content of galaxies as redshift increases. When we compare best-fit SED estimated stellar parameters for LAEs and non-LAEs, we find that Es(B-V) is smaller for LAEs at all redshifts and the difference in the median Es(B-V) between LAEs and non-LAEs increases as redshift increases, from 0.05 at z ~ 2 to 0.1 at z ~ 3.5 to 0.2 at z ~ 5. For the luminosities probed here (~L*), we find that star formation rates (SFRs) and stellar masses of galaxies, with and without Lyα in emission, show small differences such that, LAEs have lower SFRs and stellar masses compared to non-LAEs. This result could be a direct consequence of the sample selection. Our sample of LAEs are selected based on their continuum magnitudes and they probe higher continuum luminosities compared to narrow-band/emission line selected LAEs. Based on our results, it is important to note that all LAEs are not universally similar and their properties are strongly dependent on the sample selection, and/or continuum luminosities.

Non-destructive testing (NDT) is a method of testing and analysis that relies on the application of physical principles to determine the integrity of materials without causing damage. The existence of weld discontinuity can affect the service ability of the structures. Ultrasonic testing Phases Array (UTPA) is advanced ultrasonic testing technology widely practiced in many industries. The multiple ultrasonic elements and electronic time delay will create constructive and destructive interference and steering capability, which could improve detectability compared to a single element of conventional UT. The multi displays presented in A-Scan, S-scan, B-Scan, and C-Scan of UTPA equipment capable of locating, inspecting, and characterise defects within the welded component. Two carbon steel plates with thickness 18 mm and 24 mm were prepared and cut to V-shape configuration and welded using shielded metal arc welding (SMAW) process, labelled as NDE-8826 and NDE-8827 respectively. Both samples were induced with artificial defects of the weld. 16 elements of phased array probe with 0.5 mm pitch and 4 MHz were carried out for this study with encoded scanning to identify weld discontinuity plotted in different types of UT displays. Additional testing was performed using conventional ultrasonic testing (UT) using a 4 MHz probe to compare with the UTPA results and the acceptability of each defect detections. The length and datum of toe crack, slag, and lack of inter run fusion detected by UTPA in NDE-8826 give the same value as the actual value. However, the localisation of the defect is slightly different at 1.0 mm for UT detection. While the detection value for slag gives the same values for UT and UTPA, which also the same as the actual values. The detection of lack of penetration and lack of fusion in sample NDE-8827 was precisely can be detected by UT and UTPA; hence their datum value has slightly different in the centerline crack and porosity at 2.0 mm for UT measurement, and as for UTPA, it was found that the porosity at 3.0 mm which higher value compared to the actual value, 250 mm. From the result, a relevant indication from the UTPA and conventional UT collected it was found that the UTPA technique is capable of improving the Probability of Detection (POD) of defects compared to the conventional UT.

This study explored the application of nitrogen-doped graphene (NDG) ink to the coating of light-emitting diode (LED) lamps for the thermal management. Unlike the general solutions of graphene, the NDG ink used in this study was tuned with high electrical resistance and quietly suitable for the application of the LED lamp coating, which prevented the short circuit problem; besides, the NDG ink also provided a high heat dissipation effect to improve the performance of the LED lamp. The investigation of adhesion and resistance for the NDG ink were conducted by the cross-cut test and the four-point probe resistance measurement, respectively. Three types of LED lamps including the original lamp, fin-removed lamp, and NDG-ink coated (NGC) lamp were tested for the actual operating temperature distribution by K-type thermal couples and for the lumens by the integrating sphere. The results showed that the heat dissipation of the NGC lamp was better than that of the original lamp. In addition, the inspections for the properties of NDG powder were also performed by x-ray photoelectron spectroscopy (XPS), atomic force microscopy (AFM), and Raman spectroscopy. According to the inspectional results, the NDG powder had a percentage of nitrogen of about 3.8% by XPS, a surface roughness Rq of 0.9 nm by AFM, and over ten layers by Raman analysis. It showed that the NDG powder in this study belonged to the type of multi-layer graphene.

Background: The study aimed to assess whether strain ratio and T2* values can improve the sensitivity and specificity of differentiating metastatic from benign axillary lymph nodes in breast cancer patients taking histopathology as reference standard.Methods: The study was done on 43 patients. A multi-echo transverse T2*W MR sequence was obtained with TE = 0.9- 1.5 ms, TR=37.2 ms and flip angle = 25°. Sonographic elastography was done using high frequency linear probe (L3-16 MHz). Manual selection of the region of interest was done on suspicious lymph nodes for calculation of T2* values and strain ratio. ROC curves were obtained for various T2* and strain ratio values in comparison to histopathological findings as gold standard.Results: Correlation with histopathology was better with T2* values than strain ratio. The sensitivity and specificity were calculated using cut off values obtained from ROC curve (31.225 ms for T2* value and 1.85 for SR) and were 70.37%, 68.75% for strain ratio and 96.29%, 93.75% for T2* value respectively. The positive predictive value and negative predictive value were also assessed, values being higher for T2* than strain ratio. Comparison of areas under ROC curve was statistically significant with p=0.018.Conclusions: T2* can be used as a potential biomarker for differentiating metastatic from benign axillary lymph nodes owing to its high sensitivity, specificity and relative ease of performance. Quantitative assessment of changes in T2* values may allow more objective analysis of signal changes with significant differences between benign and malignant lymph nodes, even in case of partial infiltration.

Macrophage colony stimulating factor (M-CSF), a member of the group of hemopoietic cytokines, plays an important role in placental physiology of humans and mice. The objective of this study was to investigate the localization of M-CSF gene in bovine endometrial tissues during pregnancy using in situ hybridization (ISH). Twelve Japanese Black cows aged between 1.4 and 14.5 years, with normal estrous cycles, were used in this study. They were observed daily for estrous behavior, and the day of estrus was considered as Day 0. Cows were artificially inseminated, and pregnancy was confirmed by presence of conceptus and/or corpus luteum using ultrasonography. They were euthanized and their uteri were collected on Days 20 to 21 (n = 2), 36 to 42 (n = 3), 59 to 64 (n = 3), 127 to 137 (n = 2), and 225 to 226 (n = 2) of pregnancy. Pregnancy was confirmed finally by the presence of a conceptus in the uterus, and the uterus was isolated by dissection, avoiding damage of the uterine artery. The uterus was perfused with 4% paraformaldehyde (PFA) solution using catheters inserted into the uterine artery, and placental tissues were isolated by dissection. Each tissue was cut into small pieces (5 mm thick) and fixed in 4% PFA solution. After being fixed in 4% PFA solution for 20 to 24 h, tissue pieces were embedded in paraffin using routine procedures. Several tissue pieces were collected from each individual cow. Sections (6 mm thick) were cut and placed on MAS-coated glass slides (Matsunami Glass, Kishiwada, Japan). The sections were dried in an oven for 3 days at 40�C. Antisense and sense biotin-labeled oligonucleotide probes for bovine M-CSF were designed from the sequence information (GenBank accession number D87917). Sense probe was used as the negative control. ISH was carried out using the GenPoint System (DakoCytomation, Glostrup, Denmark) according to the manufacturer's instructions. The sections were observed under an Eclipse 800 microscope (Nikon, Tokyo, Japan), and the positive signal for M-CSF gene was detected. Density analysis was performed by using Scion Image (Scion Corporation, Frederick, MD, USA). Data were analyzed by one-way ANOVA and Tukey-Kramer's HSD, using 0.05 as a significant level. The M-CSF gene was expressed in the stromal and luminal epithelial cells of endometrium, stromal cells of the fetal cotyledonary villus and caruncular crypt, and mono- and multi-nuclear epithelium cells of the fetal cotyledonary villus and caruncular crypt. Intensities of M-CSF gene-positive signals in all positive cells at Days 225 to 226 of pregnancy were stronger than those at other periods (P < 0.05). These results suggest that M-CSF is produced in several cell types in the endometrium and placental tissues and may play important roles in bovine pregnancy.

Leukemia inhibitory factor (LIF), a member of the group of hemopoietic cytokines, plays a primary role in the control of embryo development and implantation and in the growth of the placenta in humans and mice. The objective of this study was to investigate the localization of LIF gene in bovine placenta tissues during pregnancy using in situ hybridization (ISH). Eleven Japanese Black cows aged between 1.8 and 14.5 years, with normal estrous cycles, were used in this study. They were observed daily for estrous behavior, and the day of estrus was considered as Day 0. They were artificially inseminated, and their uteri were collected on Days 61 to 63 (n = 3), 127 to 142 (n = 4), and 225 to 232 (n = 4) of pregnancy. Pregnancy was confirmed by the presence of a conceptus in the uterus, and the uterus was isolated by dissection, avoiding damage of the uterine artery. The uterus was perfused with 4% paraformaldehyde (PFA) solution using catheters inserted into the uterine artery, and placental tissues were isolated by dissection. Each tissue was cut into small pieces (5 mm thick) and fixed in 4% PFA solution for 20-24 h, after which they were embedded in paraffin using routine procedures. Several tissue pieces were collected from each individual cow. Six micrometer-thick sections were cut and placed on MAS coated glass slides (Matsunami Glass, Kishiwada, Japan). The sections were dried in an oven for three days at 40�C. Anti-sense and sense biotin-labeled oligonucleotide probes for bovine LIF were designed from the sequence information (GenBank accession number D50337). The sense probe was used as the negative control. ISH was carried out using GenPoint System (DakoCytomation, Glostrup, Denmark) according to the manufacturer's instructions. The sections were observed under an Eclipse 800 microscope (Nikon, Tokyo, Japan) to detect the positive signal for LIF gene. Density analysis was performed with Scion Image (Scion Corporation, Frederick, MD, USA). Data were analyzed by one-way ANOVA and Tukey-Kramer's HSD, using 0.05 as a significant level. Leukemia inhibitory factor gene was expressed in stromal cells of the fetal cotyledonary villus and caruncular crypt and in mono- and multi-nuclear epithelium cells of the fetal cotyledonary villus and caruncular crypt. Intensities of LIF gene positive signals in all positive cells at Days 127 to 142 tended to be weak compared with those at other periods. Furthermore, intensities of LIF gene positive signals of multinuclear cells of the villus and crypt demonstrated a tendency to be strong compared with those of other cells. These results suggest that LIF is produced in several cell types in the placenta and may play important roles in pregnancy.

Abstract Numerous studies have shown the prognostic importance of TP53 deletion in CLL. Assessing TP53 deletion prior to initiating therapy is good clinical practice and many laboratories now offer testing. However, there is no harmonisation of methodology and little agreement about ‘Best Practice’. Information about cut-off values is scanty in published series. The best guide to date is the LRF CLL4 trial that showed a cut-off of >20% TP53 deletion as clinically significant. This value relates to a single institution and may not be applicable to other centres using alternative sample preparation methods. We reviewed the TP53 FISH results seen in our institution and looked at the cases with levels of TP53 deletion <20% to determine their outcome. 1,424 CLL cases (Jan 2003-June 2008) were tested for deletion of TP53 using FISH (Abbott Molecular CLL probe set). Cases were referred from the Yorkshire and Humberside Cancer Network region. There were 1,020 bone marrow aspirates, 308 peripheral blood samples and 95 lymph node/tissue biopsies. CLL diagnosis was confirmed by multi-parametric flow cytometry. The criteria for FISH was the sample contained >10% B cells and had smears/dabs of suitable quality. Minor modification of standard manufacturers protocols were used to reflect the nature of the samples. Slides were examined using a Zeiss microscope and images were captured using ISIS3 (MetaSystems). Deletion analysis was microscopically scored by counting 100 lymphocytes. Normal result is defined as <5% of lymphocytes with one signal. This definition is based on our laboratory validation of the probe sets. TP53 deletion is defined as >15% of lymphocytes with one signal. Scores >5% but <15% are borderline and an additional 200 cells are scored. Results were checked by a second observer and discrepant results are referred to a third observer. In this series of CLL +12 was seen in 16.5%, mono-allelic 13q14 deletion in 43.1%, bi-allelic 13q14 deletion in 9.2%, ATM deletion in 14.2% and TP53 deletion in 11.7% of cases. Borderline results for TP53 deletion were seen in 76/1424 (5.3%) cases. No further samples were received for 38/76 (50%) cases; 18/76 (24%) were treated to CR/MRD level and CLL counts were too low for repeat FISH. 20/76 (26%) cases had additional FISH carried out, 6/20 (30%) normalised, 12/20 (60%) remained the same and 2/20 (10%) progressed to >20% TP53 deletion at 8 (25% TP53 deletion) months and 18 (>95% TP53 deletion) months. Although the technical aspects of the FISH are straightforward, and results are reproducible, the interpretation can be problematic. A small proportion of these patients show marked expansion of the TP53 clones within short time periods. Rapid clinical progression was observed in these patients. Where repeat testing was possible 2/3 had the same borderline score, these patients continue on a ‘watch & wait’ policy; the remaining 1/3 normalised their TP53 result. This suggests that there are technical quality issues with the sample. CLL cells have a tendency to disintegrate due to their fragility because of this significant subpopulations may not survive manipulation and thus evade scrutiny. A CLL patient in this series who had borderline TP53 deletion on the PB smear was found to have a TP53 level of 40% by the local referring laboratory using their cytogenetic technique. FISH on smears has an advantage over other methodologies as all CLL cells are present. There is a need for improved cross-disciplinary guidelines, and further collaborative harmonisation studies are needed. Our results show that there is no clear cut-off for a TP53 deletion result and that any result over the 5% technical cut-off needs to be reviewed. A proportion of borderline cases reflect technical difficulties of handling CLL samples but in many cases they genuinely detect a small clone of TP53 deleted cells that may expand. Careful follow-up is needed for all cases with borderline TP53 deletion results.

15508 Background: Bostwick and colleagues have suggested that prostatic intraepithelial neoplasia or PIN is the precursor of prostate cancer, and that a significant percentage of men with PIN develop prostate cancer (Sakr, 1994; Bostwick, 1995b; Arakawa, 1995; Qian, 1995; Qian, 1997). Currently there are no molecular diagnostic markers for PIN. Methods: Recently, we have developed ‘DNA-protein’ binding assays for the identification of novel transcriptional regulatory proteins associated with PIN glands. Electrophoretic mobility shift assays (EMSAs) with the 32P-labeled DNA probe was used for analysis of protein extracts from tissues and urine. Immunolabeling was carried out with rabbit antibodies specific for ABCA5. Results: Electrophoretic mobility assays (EMSAs) revealed that 1 DNA sequence (1 of 4,096 sequences screened) specifically bound a ∼160 Kda protein which was over-expressed in PIN and not found in BPH, SV or PCA glands (n=11). The gene encoding for the 160 Kda protein was cloned from a cDNA expression library by ‘in situ’ hybridization assays with the 32P-labeled DNA sequence. Sequencing revealed that the gene encoding for the 160 Kda protein was ABCA5, a member of the superfamily of ATP-binding cassette (ABC) transporters of multi-drug resistant genes. RT-PCR assays indicated the mRNA was over-expressed in PIN tissue, and was not present in BPH, or SV tissue. Rabbit polyclonal antibodies and immunolabeling showed that the antibodies specifically labeled the cell junctions of 3% glutaraldehyde fixed prostate cells. Immunohistochemistry indicated that ABCA5 was over expressed in foci of intermediate basal cells in normal glands, and in HGPIN. ABCA5 was faintly expressed in PCA glands. Enzyme linked immunoabsorbent assays (ELISAs) demonstrated in blinded studies that ABCA5 was a highly sensitive (>98% sensitivity) urine diagnostic marker for HGPIN in biopsy positive patients (n=107) at a cut off” of 25 ng/ml. ABCA5 was present at very low levels (i.e. <25 ng/ml) in the urine of patients diagnosed with BPH (n=79) or prostatitis or kidney and bladder cancer (>86% specificity). Conclusions: The data suggest that the expression of ABCA5 is over-expressed by PIN and may be a highly sensitive and specific urine diagnostic marker for detection of early stage prostate cancer. No significant financial relationships to disclose.

A simple technique was developed to fabricate carbon nanotube (CNT) probes for scanning tunneling microscope (STM). Multi-walled nanotubes were grown on the apex of the electro-chemically etched tungsten ( W ) tip using thermal chemical vapor deposition (CVD) at normal pressure with H 2 and C 2 H 2. Nickel ( Ni ) nanoparticles, which were used as the catalyst for CNT synthesis, were applied to the tip apex by dipping the W tip into the Ni nanopowder suspension in ethanol. The diameters of grown nanotubes were in the range of 20 nm to 100 nm. Their lengths were generally less than 1 μm and controlled by growth time. The technique can be readily applied to mass production of CNT STM probes without the use of any sophisticated and expensive equipments. The performance of the fabricated CNT tips was assessed by producing STM images of atomic-resolution.

Abstract Abstract 3578 Introduction: Chronic lymphocytic leukemia (CLL) patients bearing 13q14 deletion are known to experience a more favorable clinical course. Recent studies, focusing on patients with loss of 13q as the sole cytogenetic aberration at diagnosis (del13q-only cases), showed that the number of malignant cells carrying this genetic lesion correlates with a more aggressive clinical behavior. However, whether the size of the 13q deletion may also influence the clinical outcome remains to be elucidated. Patients and Methods: Probes for chromosome 13q (LSI-RB1, LSI-D13S319), 11q (LSI-ATM), 17p (LSI-p53) and chromosome 12 (CEP12) were utilized on nuclei collected at diagnosis from: i) a multi-institutional CLL cohort (342 del13q-only cases) and ii) a consecutive unselected single-institution cohort of 265 cases. RB1 deleted cases (delRB1) were defined as having at least 5% of deleted nuclei. Time to treatment (TTT) intervals, as well as Rai staging, IGHV mutational status, CD38 and ZAP70 expression, B2-microglobulin levels, all evaluated at diagnosis, were also available for all cases that entered the study. Genome wide DNA profile was performed in a pilot series of 90 CLL samples using Affymetrix GeneChip Human SNP6 arrays. Results: According to genome wide DNA analysis, delRB1 occurred in a proportion of del13q-only cases (36/90; 40%), always comprising the deleted region detected with the LSI-D13S319 probe (that covers the miR-15a/16-1 cluster and the DLEU2 gene) and characterized by a larger chromosome loss (median size 2.07 Mb vs. a median size of 0.86 Mb for the canonical del13S319). Maximally selected log-rank statistics identified the 70% of nuclei bearing del13S319 as the most appropriate cut-off value capable of separating del13q-only cases into two subgroups with different TTT distributions. Consistently, del13q-only cases with at least 70% of nuclei bearing del13S319 showed a significantly shorter TTT than del13q-only cases with less than 70% deleted nuclei (p=0.0001). Del13q-only cases were then divided in four subsets according to the percentage of nuclei bearing del13S319 with or without a concomitant delRB1: del13S319 <70% (group 1), 144 cases; del13S319 <70% + delRB1 (group 2), 95 cases; del13S319 >70% (group 3), 64 cases; del13S319 >70% + delRB1 (group 4), 39 cases. The median TTT of group 1 (not reached) was significantly longer than the median TTT of group 2 (92 months, p=0.012), group 3 (68 months, p<0.0001), and group 4 (82 months, p=0.0025; see Fig. 1A). Multivariate Cox proportional hazard analyses selected the presence of delRB1 (p=0.029), along with the IGHV mutational status (p<0.0001), as an independent negative prognosticator in the context of del13q-only cases with low/intermediate Rai risk (Rai stage of 0/I at diagnosis) and <70% of del13S319. Cases belonging to the consecutive unselected single-institution CLL cohort were divided into subsets according to the classification proposed by Döhner et al (NEJM, 2000). Notably, the presence of del13S319 in <70% of cells in the absence of delRB1 identified a patient subset with particularly stable and benign clinical course (group A in Fig. 1B, 48 cases; median TTT not reached). Conversely, patients characterized by del13S319 in <70% of cells but with a larger deletion, as determined by concomitant delRB1 (group B, 24 cases), or del13S319 in >70% of cells (with or without delRB1, group C, 25 cases) or a normal karyotype (group D, 75 cases) had shorter median TTT intervals (ranging from 105 to 129 months, p<0.01 in all the comparisons). Finally, patients affected by CLL bearing trisomy 12 (group E, 48 cases) and del11q or del17p (group F, 45 cases) experienced the worst clinical courses (p<0.0001). Conclusion: In the context of del13q-only cases, different clinical outcomes were associated to the percentage of 13q14 deleted cells, as well as to the size of the 13q14 deletion, as detected by the LSI-RB1 probe. Moreover, the presence of delRB1 emerged as a feature capable of refining the prognostic assessment in the context of CLL cases with <70% del13S319. The underlying genetic mechanisms correlated with the different clinical outcomes and associated with the size of the 13q deletion are presently under investigation. Disclosures: No relevant conflicts of interest to declare.

Nopinone based probe PPN was synthesized for the colorimetric detection of Cu²⁺ and ratiometric detection of Ag⁺, which could be applied for Ag⁺ detection in water samples and living cells.

Background: Anthrax Lethal Factor (ANT) is the dominant virulence factor produced by B. anthracis and is the major cause of death of infected animals. In this paper, pencil graphite electrodes GE were modified with single-walled and multi-walled carbon nanotubes (CNTs) for the detection of hybridization related to the ANT DNA for the first time in the literature. Methods: The electrochemical monitoring of label-free DNA hybridization related to ANT DNA was explored using both SCNT and MCNT modified PGEs with differential pulse voltammetry (DPV). The performance characteristics of ANT-DNA hybridization on disposable GEs were explored by measuring the guanine signal in terms of optimum analytical conditions; the concentration of SCNT and MCNT, the concentrations of probe and target, and also the hybridization time. Under the optimum conditions, the selectivity of probe modified electrodes was tested and the detection limit was calculated. Results: The selectivity of ANT probes immobilized onto MCNT-GEs was tested in the presence of hybridization of probe with NC no response was observed and with MM, smaller responses were observed in comparison to full-match DNA hybridization case. Even though there are unwanted substituents in the mixture samples containing both the target and NC in the ratio 1:1 and both the target and MM in the ratio 1:1, it has been found that ANT probe immobilized CNT modified graphite sensor can also select its target by resulting with 20.9% decreased response in comparison to the one measured in the case of full-match DNA hybridization case Therefore, it was concluded that the detection of direct DNA hybridization was performed by using MCNT-GEs with an acceptable selectivity. Conclusion: Disposable SCNT/MCNT modified GEs bring some important advantages to our assay including easy use, cost-effectiveness and giving a response in a shorter time compared to unmodified PGE, carbon paste electrode and glassy carbon electrode developed for electrochemical monitoring of DNA hybridization. Consequently, the detection of DNA hybridization related to the ANT DNA by MCNT modified sensors was performed by using lower CNT, probe and target concentrations, in a shorter hybridization time and resulting in a lower detection limit according to the SCNT modified sensors. In conclusion, MCNT modified sensors can yield the possibilities leading to the development of nucleic acid sensors platforms for the improvement of fast and cost-effective detection systems with respect to DNA chip technology.

Abstract The development of new anticancer agents typically involves long research cycles, high cost and low efficiency and so on. There has been a growing public interest in the use of complementary and alternative medicine for cancer treatment and drug discovery. The emergence of nanotechnology and nanomaterials has greatly stimulated research in drug delivery and optimization. As a good fluorescence probe and simultaneously as potential drug carrier, Cadmium-Tellurium quantum dots (CdTe QDs) can optimize the use and a new potential therapy method of some drug-active compounds. Based on these considerations, we have explored the possibility to connect cysteamine (Cys) modified CdTe QDs (Cys-CdTe) to gambogic acid (GA). These functional QDs were studied for multi-functional drug delivery to investigate the synergetic effect of these composites between GA and QDs for human leukemia caner cells K562 and their drug-resistant cell line K562/A02 in vitro. Also the neurotoxicity of different drug systems for Sprague Dawley (SD) rat brain was analyzed by in vivo real-time microdialysis. The nanocomposites of GA loaded CdTe QDs were prepared and the drug releasing of GA-CdTe nanocomposites was performed at the medium of pH 6.0 and 7.4 in vitro. The cytotoxicity of GA and the nanocomposites for K562 and K562/A02 cells was studied by MTT, cell morphology, and flow cytometry. The labeling and imaging of CdTe QDs and the nanocomposites for cancer cells was performed by laser confocal fluorescence microscopy. Different drug systems were respectively injected into SD rat by vena caudalis and cerebrospinal fluid was collected by microdialysis in vivo. The neurotoxicity of different drug systems for rat brain was investigated by the changes of amino acid content in the cerebrospinal fluid. The Cys modified CdTe QDs showed the good fluorescence characteristics, with the wide excitation spectrum and narrow emission spectrum. The CdTe QDs could label in human leukemia cancer cells not only for K562 but also K562/A02 , which had the potential application in the cell imaging. Following the electrostatic attraction, surface adsorption et al, these CdTe QDs had the good capability of drug delivery with the high drug-loading and envelopment capacity. Also these GA-CdTe nanocomposites showed the pH sensitivity for the drug release and improved the drug's un-dissolvability. The GA-CdTe nanocomposites also had the fluorescence characteristics for labeling the different kinds of leukemia cancer cells. Gambogic acid play an important role in cancer therapy and also showed the sensitivity for the multidrug resistance (MDR) of K562/A02 cells in this study. The results indicated that GA-CdTe nanocomposites could significantly enhance the drug accumulation to improve the cytotoxicity and considerably overcome the multidrug resistance. GA-CdTe nanocomposites induced the arrest of G0/G1 phase to promote the cells apoptosis. The combination between GA and CdTe QDs can optimize the new potential therapeutic method for GA with the real time labeling and tracing during the disease therapy. The neurotoxicity of GA and GA-CdTe nanocomposites for rat brain was firstly explored by using the microdialysis in vivo. The results indicated that GA had the serious neurotoxicity through the concentration changes of amino acids for the rat brain. The concentration of some excitatory amino acids was remarkably increased. For the GA-CdTe nanocomposites based on CdTe QDs, the side effect of GA was visibly cut down, and the time to cause the neurotoxicity was apparently shortened. These nanocomposites own the better biocompatibility and bio-safety for the relevant cancer treatment in vitro and in vivo. This raises the promising possibility of the application of these fluorescent nanocomposites based on CdTe QDs for the target cancer therapy. The combination of GA with Cys-CdTe QDs can optimize the use and new potential multi-mode therapy of cancers. This may shed new light to exploit the potential application of the active compounds from Traditional Chinese Medicine (TCM). Disclosures: No relevant conflicts of interest to declare.

Although agonistic interactions between cats are often regarded clinically as a source of stress, there is currently limited research evidence regarding the welfare impact of keeping multiple cats as pets. The aim of this study was to compare welfare indicators between cats living in domestic single and multi-cat households, as well as between multi-cat households where agonistic behaviour was/was not reported by owners. Indicators included a spatial judgment bias task (JBT), where longer latencies to ambiguous probes are interpreted as being related to a more ‘pessimistic’ mood state, and the cat stress score (CSS), where high scores are indicative of high stress levels. Of 128 focal cats between the ages of 9–22 months, 94 were from multi-cat households, 126 had useable CSS data and 42 had JBT results suitable for analysis. CSSs were significantly lower for cats showing a more ‘pessimistic’ response in the JBT. It is possible that the cats that appeared to be the most relaxed may have been showing inactivity relating to negative affective states and/or were the least active/food motivated, and therefore slower in the JBT. CSSs were significantly higher in cats from single compared with multi-cat households, and did not vary with reports of agonistic interactions in multi-cat households. JBT results did not vary depending on the presence of, or reports of agonistic behaviours between, cohabiting cats. These data suggest that cats from single-cat households may be more likely to show signs of acute stress than those in multi-cat households. Alternative explanations are possible. For example, lower CSSs in the multi-cat group may reflect ‘relief’ effects resulting from separating cats for the test period, or inactivity relating to negative affective states. Due to the narrow sample population and broad scope of husbandry conditions, the potential for confounding variables limits the degree by which results can be used to inform causation of the relationships identified. Further research is warranted to replicate this work and explore potential confounders.

A novel electrochemical DNA biosensor system based on multi-walled carbon nanotubes(MWCNTs) was presented in this paper. In order to make multi-walled carbon nanotubes being perpendicularly standing on Au electrode via Au-S chemical bonding, we cut multi-walled carbon nanotubes (1.5 m) into short pipes (200-300 nm). Then, the shortened nanotubes were further modified by thiolization reaction with cysteamine (NH2CH2CH2SH). The –SH modified MWCNTs, gold nanoparticles and 5’ end –SH modified DNA (HS-DNA) as nucleotide probes were self-assembled onto the surface of Au electrode respectively. The DNA biosensors based on self-assembled MWCNTs had a higher current response compared to those based on random MWCNTs. When the target DNA concentration was 4.0×10–9 mol/L to 1.2×10-8 mol/L, the cathodic peak current of Au electrode system with the AQDS as indicator was linearly related, and the detection limit was about 6.5×10-9 mol/L. In addition, the developed DNA biosensors also had a high selectivity of hybridization detection.

Recently intensive research has been carried out on carbon nanotube (CNT) based polymer composites. However, in this work the macro scale IM700 carbon fiber (CF) has been integrated with amino modified multi-wall carbon nanotubes (MWCNT-NH2) within LY-556 epoxy matrix to produce three-phase, multi-scale composites, with the applications in missiles. The fictionalization of CNTs has been carried out for improved and consistent mechanical and physical properties. High frequency probe sonication method was used for homogenous dispersion of CNTs. Mechanical characterization of the multiscale composites fabricated by hand layup process included tensile, flexure and inter-laminar shear stress tests. The addition of small amounts of MWCNTs (upto 1.5 weight %) for the fabrication of multiscale composites resulted in a maximum enhancement in tensile strength by 23% , flexural modulus by 35%, flexural strength by 5% and ILSS by 7%. CF/epoxy composites (without CNTs) have also been characterized for comparison with MWCNT-NH2/CF/ epoxy composites. For a more accurate prediction of the Young’s moduli of multi-scale composites several micromechanical models (Voigt-Reuss, Halpin-Tsai, and modified mixture Law) compared with the experimental work. These models have been tested for upto 1.5 weight % of CNTs. The difference of approximate 28% of the values in Young’s modulus has been reported in the classical micromechanical models and experimental results and it comes out 11% for CF/epoxy composites as they do not have reinforcement of CNT’s within it. In all the classical models, the Young’s moduli of the carbon nanotube composites were used as matrix properties.

Current soil classification systems based on cone penetration testing (CPT) utilize a combination of the tip resistance (qt), pore pressure (u2), and friction sleeve (fs) measurements as inputs. While the qt measurements are typically normalized by the overburden stress, the fs measurements are often normalized by the net tip resistance, leading to the use of parameters that are dependent on each other. This paper presents the development of a soil classification framework that utilizes a normalized multi-friction parameter (MFP) and the CPT normalized tip resistance. The MFP parameter is obtained from measurements with textured friction sleeves from soundings with multi-sleeve attachments. The use of textured friction sleeves allows for fundamental differences in soil–structure interface behavior and particle sizes to be captured due to the significant degree of shearing induced within the soil. This classification framework was developed with results from over 30 soundings at six different sites. The analysis of samples taken from the field indicates that the proposed framework provides a classification that better agrees with the grain-size distribution for residuum, calcareous and intermediate soils, as compared to existing CPT-based systems. The potential development of a simplified probe with just one additional friction sleeve sensor can provide appropriate classification results and would facilitate adoption for use in practice.

Cet article se fonde sur l’intuition que l’on peut appréhender les pratiques vidéoludiques contemporaines, particulièrement pour les jeux de rôle en lignes massivement multi-joueurs, à partir de ce rapprochement que pose Hamayon entre, jeu, logique de chasse, anticipation et gestion de l’aléatoire. Après un état des lieux des logiques de la chasse dans l’univers deWorld of Warcraftsont décrites les stratégies cynégétiques des joueurs. Sur cette base est clarifiée la dialectique proie-prédation qui charpente les relations entre joueurs et non-humains informatiques dans ce monde numérique. Échappe à ces relations habituelles la figure du prédateur absolu, impossible à abattre seul, et qui fait que le chasseur peut se retrouver chassé. Les techniques d’anticipation collective, notamment en guilde, deviennent une des nouvelles ressources de survie, indispensables surtout pour gérer la part aléatoire de ce type de confrontation. C’est à ces stratégies de chasse particulières et à ces techniques de prévision que s’intéresse le dernier volet de cette analyse, y compris la gestion du butin aléatoire issu de la victoire collective contre de tels monstres digitaux.

The main raw material of solar energy is multi-crystalline silicon. Directional solidification technique is one important technological process of metallurgy purification technology for multi-crystalline silicon. It can purify metallurgical grade silicon by removing metal impurities and control crystal growth at the same time. In experiment, metallurgical grade silicon by acid leaching pre-treatment, was purified by our self-assembled directional solidification furnace. The sample was analyzed by electron-prode micro analysis (EPMA). According to the results, the removal efficiency of Fe and Al is 96.3% and 96.7%, respectively. The removing mechanism of metal impurities and the difference between theory value and experiment value were also discussed. The segregation effect in directional solidification is the reason of removing Fe, but analgesic effects of the segregation effect combined with vacuum volatilization are that of removing Al. When the silicon ingot was cooled down, lengthways section of silicon ingot was cut and etched, crystal growth was studied. The results indicate that columnar crystal growth shows diverging tendency from the bottom to the top of silicon ingots, and solidification interface shape is convex. The reasons may be the nucleation of new crystals on crucible sidewall is very serious and the pulling rate is too high.

The microbial fuel cell (MFC) is a novel and innovative technology that could allow direct harvesting of energy from wastewater through microbial activity with simultaneous oxidation of organic matter in wastewater. Among all MFC parts, electrode materials play a crucial role in electricity generation. A variety of electrode materials have been used, including plain graphite, carbon paper and carbon cloth. However, these electrode materials generated only limited electricity or power. Recently, many research studies have been conducted on carbon nanotubes (CNTs) because of their unique physical and chemical properties that include high conductivity, high surface area, corrosion resistance, and electrochemical stability. These properties make them extremely attractive for fabricating electrodes and catalyst supports. In this study, CNT-based electrodes had been developed to improve MFC performance in terms of electricity generation and treatment efficiency. Multi-walled carbon nanotubes (MWCNTs) with carboxyl groups have been employed to fabricate electrodes for single-chamber air-cathode MFCs. The quality of the prepared MWCNTs-based electrodes was evaluated by morphology, electrical conductivity and specific surface area using a field emission scanning electron microscope, four-probe method and Brunauer–Emmerr–Teller method, respectively. The performance of MFCs equipped with MWCNT-based electrodes was evaluated by chemical analysis and electrical monitoring and calculation. In addition, the performance of these MFCs, using MWCNTs as electrodes, was compared against that using commercial carbon cloth.

Abstract. We present an innovative instrument telescope, and describe a retrieval method to probe 3-D distributions of atmospheric trace gases that are relevant to air pollution and tropospheric chemistry. The University of Colorado (CU) two dimensional (2-D) Multi-AXis-Differential Optical Absorption Spectroscopy (CU 2D-MAX-DOAS) instrument measures nitrogen dioxide (NO2), formaldehyde (HCHO), glyoxal (CHOCHO), oxygen dimer (O2-O2, or O4) and water vapor (H2O); also nitrous acid (HONO), bromine monoxide (BrO), iodine monoxide (IO) among other gases can in principle be measured. Information about aerosols is derived through coupling with a radiative transfer model (RTM). The 2-D telescope has 3 modes of operation: (mode 1) measures solar scattered photons from any pair of elevation angle (−20° < EA < +90° or zenith; zero is to the horizon) and azimuth angle (−180° < AA < +180°; zero being North), (mode 2) measures any set of AA at constant EA (almucantar scans); and (mode 3) tracks the direct solar beam via a separate view port. Vertical profiles of trace gases are measured, and used to estimate planetary boundary layer height (PBL). Horizontal distributions are then derived using PBL and parameterization of RTM (Sinreich et al., 2013). NO2 is evaluated at different wavelengths (350, 450, and 560 nm), exploiting the fact that the effective path length varies systematically with wavelength. The area probed is constrained by O4 observations at nearby wavelengths, and has an effective radius of 7.5 to 20 km around the instrument location; i.e., up to 1250 km2 can be sampled near-instantaneously, and with high time resolution. The instrument was deployed as part of the Multi Axis DOAS Comparison campaign for Aerosols and Trace gases (MAD-CAT) in Mainz, Germany from 7 June to 6 July 2013. We present first measurements (modes 1 and 2 only) and describe a four-step retrieval to derive (a) boundary layer vertical profiles of NO2 and PBL; (b) near-surface horizontal distributions of NO2; (c) range resolved NO2 horizontal distribution measurements using an "onion peeling" approach; and (d) the ratios HCHO-to-NO2 (RFN), CHOCHO-to-NO2 (RGN), and CHOCHO-to-HCHO (RGF) at 14 pre-set azimuth angles distributed over a 360° view. 2D-MAX-DOAS provides an innovative, regional perspective about trace gases, their spatial and temporal concentration gradients, and maximizes information to compare near-surface observations with atmospheric models and satellites.

Natural herbicides that are based on allelopathy of compounds, can offer effective alternatives to chemical herbicides towards sustainable agricultural practices. Nerolidol, a sesquiterpenoid alcohol synthesized by many plant families, was shown to be the most effective allelopathic compound in a preliminary screening performed with several other sesquiterpenoids. In the present study, Arabidopsis thaliana seedlings were treated for 14 d with various cis-nerolidol concentrations (0, 50, 100, 200, 400, and 800 µM) to investigate its effects on root growth and morphology. To probe the underlying changes in root metabolome, we conducted untargeted gas chromatography mass spectrometry (GC-MS) based metabolomics to find out the specificity or multi-target action of this sesquiterpenoid alcohol. Oxidative stress (measured as levels of H2O2 and malondialdehyde (MDA) by-product) and antioxidant enzyme activities, i.e., superoxide dismutase (SOD) and catalase (CAT) were also evaluated in the roots. Nerolidol showed an IC50 (120 µM), which can be considered low for natural products. Nerolidol caused alterations in root morphology, brought changes in auxin balance, induced changes in sugar, amino acid, and carboxylic acid profiles, and increased the levels of H2O2 and MDA in root tissues in a dose-dependent manner. Several metabolomic-scale changes induced by nerolidol support the multi-target action of nerolidol, which is a positive feature for a botanical herbicide. Though it warrants further mechanistic investigation, nerolidol is a promising compound for developing a new natural herbicide.

Cone penetration test is a fast and efficient in-situ test technique. With the development of sensor technology and the use of new probes, such test is employed in more fields and reveals more soil parameters. Based on advanced CPTU equipment, soil types were classified. As CPTU has the function of porewater pressure test, the value of porewater pressure varies a lot when the soil changes, which is shown clearly in the CPTU feature map. So it can be easier to judge soil boundaries and its result is in good agreement with the borehole. Multi-function CPTU system is equip with SCPTU module, which enable to measure shear wave velocity of the soil easily. To meet the needs of conventional CPT equipment, the relationships between shear wave velocity measured by SCPTU and other CPT indexes were analyzed; then, two empirical formulas which suitable for kinds of soils are proved more consistent with the measured results, so it is a good method to estimate shear wave velocity without seismic wave test. With a view to get greater economic and technical benefits, more cone penetration testing experience in different regions should be accumulated for geotechnical engineering investigation and design.

When presented with simple stimuli like bars and gratings, adjacent neurons in striate cortex exhibit shared selectivity for multiple stimulus dimensions, such as orientation, direction, and spatial frequency. This has led to the idea that local averaging of neuronal responses provides a more reliable representation of stimulus properties. However, when stimulated with complex, time-varying natural scenes (i.e., movies), striate neurons exhibit highly sparse responses. This raises the question of how much response heterogeneity the local population exhibits when stimulated with movies, and how it varies with separation distance between cells. We investigated this question by simultaneously recording the responses of groups of neurons in cat striate cortex to the repeated presentation of movies using silicon probes in a multi-tetrode configuration. We found, first, that the responses of striate neurons to movies are brief (tens of milliseconds), decorrelated, and exhibit high population sparseness. Second, we found that adjacent neurons differed significantly in their peak firing rates even when they responded to the same frames of a movie. Third, pairs of adjacent neurons recorded on the same tetrodes exhibited as much heterogeneity in their responses as pairs recorded by different tetrodes. These findings demonstrate that complex natural scenes evoke highly heterogeneous responses within local populations, suggesting that response redundancy in a cortical column is substantially lower than previously thought.

Microorganisms have begun to develop resistance because of inappropriate and extensive use of antibiotics in the hospital setting. Therefore, it seems to be necessary to find a way to tackle these pathogens by developing new and effective antimicrobial agents. Carbon nanotubes (CNTs) have attracted growing attention because of their remarkable mechanical strength, electrical properties, and chemical and thermal stability for their potential applications in the field of biomedical as therapeutic and diagnostic nanotools. However, the impact of carbon nanotubes on microbial growth has not been fully investigated. The primary purpose of this research study is to investigate the antimicrobial activity of CNTs, particularly double-walled and multi-walled nanotubes on representative pathogenic strains such as Gram-positive bacteria Staphylococcus aureus, Gram-negative bacteria Pseudomonas aeruginosa, Klebsiella pneumoniae, and fungal strain Candida albicans. The dispersion ability of CNT types (double-walled and multi-walled) treated with a surfactant such as sodium dodecyl-benzenesulfonate (SDBS) and their impact on the microbial growth inhibition were also examined. A stock concentration 0.2 mg/mL of both double-walled and multi-walled CNTs was prepared homogenized by dispersing in surfactant solution by using probe sonication. UV-vis absorbance, Fourier transform infrared spectroscopy (FTIR), and transmission electron microscopy (TEM) were used for the characterization of CNTs dispersed in the surfactant solution to study the interaction between molecules of surfactant and CNTs. Later, scanning electron microscopy (SEM) was used to investigate how CNTs interact with the microbial cells. The antimicrobial activity was determined by analyzing optical density growth curves and viable cell count. This study revealed that microbial growth inhibited by non-covalently dispersed CNTs was both depend on the concentration and treatment time. In conclusion, the binding of surfactant molecules to the surface of CNTs increases its ability to disperse in aqueous solution. Non-covalent method of CNTs dispersion preserved their structure and increased microbial growth inhibition as a result. Multi-walled CNTs exhibited higher antimicrobial activity compared to double-walled CNTs against selected pathogens.

Abstract. We present an innovative instrument telescope and describe a retrieval method to probe three-dimensional (3-D) distributions of atmospheric trace gases that are relevant to air pollution and tropospheric chemistry. The University of Colorado (CU) two-dimensional (2-D) multi-axis differential optical absorption spectroscopy (CU 2-D-MAX-DOAS) instrument measures nitrogen dioxide (NO2), formaldehyde (HCHO), glyoxal (CHOCHO), oxygen dimer (O2–O2, or O4), and water vapor (H2O); nitrous acid (HONO), bromine monoxide (BrO), and iodine monoxide (IO) are among other gases that can in principle be measured. Information about aerosols is derived through coupling with a radiative transfer model (RTM). The 2-D telescope has three modes of operation: mode 1 measures solar scattered photons from any pair of elevation angle (−20° < EA < +90° or zenith; zero is to the horizon) and azimuth angle (−180° < AA < +180°; zero being north); mode 2 measures any set of azimuth angles (AAs) at constant elevation angle (EA) (almucantar scans); and mode 3 tracks the direct solar beam via a separate view port. Vertical profiles of trace gases are measured and used to estimate mixing layer height (MLH). Horizontal distributions are then derived using MLH and parameterization of RTM (Sinreich et al., 2013). NO2 is evaluated at different wavelengths (350, 450, and 560 nm), exploiting the fact that the effective path length varies systematically with wavelength. The area probed is constrained by O4 observations at nearby wavelengths and has a diurnal mean effective radius of 7.0 to 25 km around the instrument location; i.e., up to 1960 km2 can be sampled with high time resolution. The instrument was deployed as part of the Multi-Axis DOAS Comparison campaign for Aerosols and Trace gases (MAD-CAT) in Mainz, Germany, from 7 June to 6 July 2013. We present first measurements (modes 1 and 2 only) and describe a four-step retrieval to derive (a) boundary layer vertical profiles and MLH of NO2; (b) near-surface horizontal distributions of NO2; (c) range-resolved NO2 horizontal distribution measurements using an "onion-peeling" approach; and (d) the ratios HCHO to NO2 (RFN), CHOCHO to NO2 (RGN), and CHOCHO to HCHO (RGF) at 14 pre-set azimuth angles distributed over a 360° view. Three-dimensional distribution measurements with 2-D-MAX-DOAS provide an innovative, regional perspective of trace gases as well as their spatial and temporal concentration gradients, and they maximize information to compare near-surface observations with atmospheric models and satellites.

Field emission properties of individual multi-walled carbon nanotubes (MWNTs) were studied in nanorobotic manipulation and electron-beam-induced deposition (EBID). Nanotube emitters are constructed by picking up and assembling individual nanotubes on a commercially available atomic force microscope (AFM) cantilever or a tungsten probe. The relationship between field emission current and interelectrode distance was obtained by changing the distance between the tip of the nanotube emitter and the counterpart anode, which can be potentially applied as the principle for an approaching sensor to detect nanometer scale distance by observing field emission current in real time. Field emission current on a microampere scale from a CNT emitter was shown to be strong enough for EBID without obviously degrading emitters. Deposit topology was related to current density or the emitter shape, suggesting that information on emitter geometry could be obtained from EBID deposits. Energy dispersive X-ray spectrometry (EDS) analysis of deposits from W(CO)6showed that the tungsten mass exceeds 80% on the average among compositions. Much higher voltage may degrade the emitter, and saturated current may be used to adjust the emitter length in a controlled way.

Abstract The paper presents a novel antenna operating at the lower UWB band (3.75–4.25 GHz), defined originally in IEEE 802.15.6 standard for Body Area Networks (BAN) applications. The proposed antenna is designed for biomedical application, wireless capsule endoscopy localization. In other words, the concerned application is dedicated to track a capsule, by means of an external device, swallowed by the patient to provide captured images of the Small Intestine (SI), essential part of the GastroIntestinal (GI) tract, and transfer them in real-time to the external device. In this context, antenna with and without cavity-backed structures, are presented and compared with the requirements for a receiving antenna in terms of directivity and bandwidth coverage in question. It was revealed that the cavity approach improved the antenna gain up to 8 dBi, at the 4 GHz center frequency, compared to 6 dBi without the cavity presence. Simulations were carried out using CST Microwave Studio, and the results were validated by measurements in proximity to human body. The antenna safety issue was assessed with CST SAR (Specific Absorption Rate) calculation, in compliance with IEEE/IEC 62704-1 standard. Results showed a maximum SAR of 0.112 W/kg and 0.005 W/kg at 4 mm and 30 mm antenna-skin distance, in the range of the SAR limit guidelines defined by safety standards. The cavity-backed antenna ability to penetrate the human tissues, to reach the small intestine layer was studied by means of CST voxel model and compared to a multi-layer model emulating the dielectric properties of the human tissues at 4 GHz. This analysis was conducted using power flow results and completed by the power field probes at the several tissue interfaces.

AbstractX- and Y-mediated input to areas 17 and 18 of the cat visual cortex was studied using current-source-density analysis of field potentials evoked by stimulation of the optic nerves. A cuff-shaped electrode was used for stimulation so that Y axons, by virtue of their larger diameters, would have lower electrical thresholds than X axons. The effect in each cortical area of activating Y axons alone could therefore be determined by low-&amp;litude stimulation of the optic nerves. Current-source densities were calculated by two separate methods. (1) In five experiments, field potentials were measured sequentially at different cortical depths with a single tungsten electrode. Current densities were then calculated by computer. (2) In two experiments, current densities were derived in real time from field potentials recorded simultaneously from three sites with a multi-electrode probe. The calculation was performed by an analog circuit specially designed for this purpose. This method has several advantages over the standard, single-electrode method. At stimulus strengths sufficient to activate the majority of Y axons in the optic nerves, but subthreshold to most X axons, the field potentials evoked in area 17 changed little from layer to layer. When the current-source-density analysis was applied to these potentials, no significant sources or sinks were detectable. Only when the stimulus strength was raised to the point that both X and Y axons were activated by the stimulus were any current sources or sinks detected in area 17. The currents were similar in time course and laminar pattern to those recorded after stimulation of the optic chiasm. In area 18, large sources and sinks were evoked by stimulation of Y axons alone. These currents changed little when the stimulus strength was increased to activate X axons as well. Area 18, therefore, in contrast to area 17, seems to be dominated by Y input and receives little X input. These results support the conclusions of the accompanying paper in which synaptic potentials were recorded intracellularly from cortical neutrons. The intracellular experiments failed to show substantial Y input to area 17. The projections of X and Y axons may therefore be much more highly segregated into areas 17 and 18 than previously thought. Alternatively, the nature of the Y input to area 17 may be very different from that to area 18 in that it cannot be easily detected with intracellular or current-source-density techniques.

AbstractSulphur isotopes can be used as a powerful tool to trace fluid evolution and explore the formation of chimneys. To clarify the in situ S isotopic variations of sulphides at the micro-scale, we analyzed a sulphide chimney collected from the hydrothermal field in the East Pacific Rise 1–2° S using a sensitive high-mass-resolution ion micro-probe for stable isotopes (SHRIMP SI). Three mineral zones can be identified in the chimney: an external outer wall of porous anhydrite and colloform pyrite, an internal middle zone of sub-euhedral pyrite and massive chalcopyrite, and an inner zone of massive pyrite. The δ34SV-CDT values of the sulphides fall within the range 1.83–7.51 ‰ (avg. 4.05 ‰, n = 16), and S isotopic values increase from the core (3.09 ‰, n = 3) to the middle (3.78 ‰, n = 11) to the edge (6.99 ‰, n = 2). These results illustrate mineral crystallization processes and the mixing between seawater-derived S and magmatic–hydrothermal fluids during the growth of the chimney. The zones from the edge to the core are characterized by crystal morphologies of colloform/anhedral pyrite to massive pyrite with decreasing δ34S values, revealing multi-stage mineral deposition and sulphur isotopic fractionation. In contrast to the increase in δ34S values from the core to the edge in one profile (profile A), anomalously low δ34S values in fine-grained pyrite relative to chalcopyrite in another profile (profile B) in the middle zone result from S isotopic exchange between seawater SO42− and fluid H2S due to different fluid–seawater mixing, possibly caused by variations in permeability and porosity across the chimney.

Extragalactic planetary nebulae (PNe) are detectable through relatively strong nebulous [O III] emission and act as direct probes into the local stellar population. Because they have an apparently universal invariant magnitude cut-off, PNe are also considered to be a remarkable standard candle for distance estimation. Through detecting PNe within the galaxies, we aim to connect the relative abundances of PNe to the properties of their host galaxy stellar population. By removing the stellar background components from FCC 167 and FCC 219, we aim to produce PN luminosity functions (PNLF) of these galaxies, and thereby also estimate the distance modulus to these two systems. Finally, we test the reliability and robustness of our novel detection and analysis method. It detects unresolved point sources by their [O III] 5007 Å emission within regions that have previously been unexplored. We model the [O III] emissions in the spatial and spectral dimensions together, as afforded to us by the Multi Unit Spectroscopic Explorer, and we draw on data gathered as part of the Fornax3D survey. For each source, we inspect the properties of the nebular emission lines to remove other sources that might hinder the safe construction of the PNLF, such as supernova remnants and H II regions. As a further step, we characterise any potential limitations and draw conclusions about the reliability of our modelling approach through a set of simulations. By applying this novel detection and modelling approach to integral field unit observations, we report for the distance estimates and luminosity-specific PNe frequency values for the two galaxies. Furthermore, we include an overview of source contamination, galaxy differences, and possible effects on the PNe populations in the dense stellar environments.

A slaughterhouse study commissioned by Florida Cattleman’s Association in 2007 identified ovarian follicular dysplasia (OFD) as a primary cause of infertility in Florida beef cows. Ovaries with OFD have progressive bilateral development of solid clustered follicles containing multiple Call-Exner bodies that originate in the rete ovarii and the hilar region, and progress into the cortex to eventually form bilateral Sertoli-type granulosa theca cell tumours (GTCT). The objectives of this study were to assess the distribution of OFD in cull animals and to evaluate utilisation of ultrasound for diagnosis of OFD in cattle. Ultrasound images of the right and left ovaries from 390 cull cows and heifers representing 4 Florida ranches were made with 5-MHz linear probes (Aloka, Ibex). Then, 10 to 12 females per ranch were followed to slaughter the proceeding day for collection of reproductive tracts. The fixed ovaries were measured, sectioned para-sagittally through the hilus, photographed, and arranged in histology cassettes for complete examination of the cut surface. Large ovarian structures including corpus luteum, Graafian follicles, atretic follicles, dysplastic follicles, rete ovarii, dysplastic follicles, and tumours were counted and measured for each ovary. Ovaries with OFD were graded I to IV. Grade I OFD contained small individual dysplastic follicles with diameter less than 200 µm mostly limited to the rete ovarii and medulla. Grade II OFD possessed dysplastic follicles greater than 200 µm diameter that were present in the medulla and cortex. Grade III OFD had extensive multi-sized dysplastic follicles scattered throughout the entire cortex of the ovary and Grade IV OFD had Sertoli-type GTCT. Grade II–IV often had dystrophic mineralization of dysplastic follicles. Gross morphology of fixed sagittal sections and ultrasound images were blindly compared against OFD grade in 40 individual ovaries. The OFD was identified at slaughter in 29/41 cows and in 1/5 of heifers. The distribution of OFD for 30 affected females was Gr I 16/30, Gr II 9/30, Gr III 4/30, and Gr IV 1/30. Characteristics that could be detected by routine ultrasound included increased size and length, increased hyperechogenicity and decreased number of fluid-filled follicles. Hyperechogenic shadows were evident in higher grade OFD. The study demonstrated that Grade III and IV OFD can be observed by routine ultrasound but Grade I and II may require higher resolution ultrasound probes, imaging analysis software, or Doppler ultrasound.

Abstract. Despite all the attention paid to the topic of policy analysis as a conceptual endeavour, empirically, the actual work of policy analysts is little investigated and little known. This is true generally of most countries and jurisdictions but it is most acute at the subnational level of government in multilevel states. Recent work in Canada, however, based on comprehensive surveys of analysts of provincial and territorial policy, on the one hand, and regionally and Ottawa-based federal policy workers on the other, has found many similarities with national-level work but also significant differences. This work has highlighted differences in the distribution of tasks across jurisdictions—mainly the extent to which policy work involves implementation as well as formulation-related activities—as key distinctions found in policy work across levels of the Canadian multilevel system. This article uses frequency and principal components analysis (PCA) and structural equation modeling (SEM) to probe these dimensions of policy work. It shows provincial and territorial analysts to be similar to regionally based federal workers in task allocation, undermining a straightforward depiction of differences in policy work by level of government. The extent of autonomy enjoyed by policy workers in different jurisdictional venues, both from internal actors and those outside of government, is shown to be the key driver of differences in policy work across levels of government.Résumé. Malgré toute l'attention accordée au thème de l'analyse politique comme un effort conceptuel, empirique du travail réel des analystes des politiques est peu étudié et mal connu. Ceci est vrai en général de la plupart des pays et juridictions, mais est le plus aigu au niveau sous-national de gouvernement dans les États multi-niveaux. Des travaux récents au Canada, cependant, basée sur des enquêtes complètes des provinces et des territoires, d'une part, et régional et basée à Ottawa analystes de la politique fédérale, d'autre part, a trouvé de nombreuses similitudes avec le travail au niveau national mais aussi des différences significatives. Ce travail a mis en évidence des différences dans la répartition des tâches entre les administrations – notamment la mesure dans laquelle le travail politique consiste à la mise en œuvre ainsi que la formulation des activités liées – comme les distinctions clés trouvés dans le travail politique à travers les niveaux de l'canadienne système multi-niveau. Cet article utilise la fréquence et analyse en composantes principales (ACP) et la modélisation par équations structurelles (SEM) pour sonder ces dimensions du travail politique. Il montre les analystes provinciaux et territoriaux à être semblables à l'échelle régionale basée sur les travailleurs fédéraux dans la répartition des tâches, minant une représentation directe des différences dans le travail politique, par niveau de gouvernement. Le degré d'autonomie dont jouissent les travailleurs dans les différents lieux de la politique juridictionnelle – à la fois par des acteurs internes et ceux de l'extérieur du gouvernement – se révèle être le principal moteur de différences dans le travail politique à travers les niveaux de gouvernement.

The development of methods for qualitative and quantitative analysis of drugs can guarantee their identity and quality. Drugs used in the form of salts of organic bases are often determined by the anionic part of these salts. Citrate ions are a part of many drugs in the form of citric acid, salts of alkaline and alkaline earth metals. The purpose of this study was to develop a method for the luminescent determination of citrate ions in dosage forms using a complex of yttrium (III) with rutine (Rut) as a luminescent probe. It has been experimentally established that citrate ions increase the luminescence intensity of the Y(III)–Rut complex. The spectral and luminescence characteristics of the complex was studied. The luminescence spectrum of the Y(III)–Rut complex has a maximum at λlum = 570 nm. The luminescence intensity of the Y(III)–Rut complex increases and the luminescence peak shifts to the short-wave region of the spectrum (λlum = 522 nm) in the presence of sodium citrate. The maximum effect is observed at a pH of 6.5–7.5. The dependencies of the luminescence intensity on the concentration of Y(III) and Rut for the Y(III)–Rut–Cit complex at the constant concentration of citrate ions (1·10-3 mol/l) were studied. It was established that the maximum luminescence intensity was observed at concentrations of Y(III) – 2·10-3 mol/l and Rut – 5·10-4 mol/l. The linear region of the dependence of the luminescence intensity of the complex on the concentrations of Y(III) and Rut is observed in the range of yttrium concentrations 0.3–2.0·10-3 mol/l and rutine 0.5–5.0·10-4 mol/l. The method of luminescent determination of citrate ions in dosage forms has been developed. The method is based on the use of rutine molecular luminescence in the multi-ligand complex Y(III)–Rut–Cit. The method of determination of citrate ions in dosage forms differs favorably from the existing absence of toxic reagents, expensive equipment, short-term analysis time, allows rapid screening of samples of drugs.

Welding is one of the most versatile joining methods for constructing products and structures in nearly all industrial fields. Arc has been widely used as a cheap heat source for welding since carbon arc fusion welding was first applied to join Pb plates in about 1880. New welding technologies have been developed according to social needs or changes since 1960. Therefore, half-automated welding, automatic welding and highefficient welding have been developed for saving man-power and afterward full automation. First, tandem one-side SAW (submerged arc welding), high-speed rotational arc, high-heat input SAW, tandem wire MAG, etc. have been introduced as highly efficient welding processes. On the other hand, as gas-shielding arc welding processes, CO2 gas, MAG, man-power saving automatic welding, the use of a flux-cored wire, AC MIG, MIG with two wires, laser-arc hybrid welding, CMT process have been developed and most widely employed in the industries in conjunction with an advance in the welding heat sources from thyristor to inverter and nowadays digital inverter. Furthermore, robotization has been developed from spot welding robot to squire robot, multi-axes GAM robot, mobile robot, portable many-axes robot and 7 axes robot together with the development in welding sensors such as probe sensor, one-touch sensor, magnetic sensor, arc sensor, laser-slit light sensor, stereo CCD, etc. Recently, novel arc sources are not developed, but deep weld penetration and geometry are controllably obtained in TIG welding by active flux pasted on the plate surface, good use of an active gas and narrow oxidation treatment. Clean MIG process for steels is also developed by use of a unique solid-wire of double layers with different melting temperatures, and different hybrid heat sources of plasma and GMA or laser and MIG. Hybrid welding processes with CO2 laser and MAG, disk laser and MAG, fiber laser and CO2 arc or MAG has recently been applied in the shipbuilding industry. I thank the authors for their generous cooperation to the publication of new development in the welding technologies.

Abstract. New particle formation (NPF), which generates nucleation mode aerosol, was observed in the tropical Upper Troposphere (UT) and Tropical Tropopause Layer (TTL) by in situ airborne measurements over South America (January–March 2005), Australia (November–December 2005), West Africa (August 2006) and Central America (2004–2007). Particularly intense NPF was found at the bottom of the TTL. Measurements with a set of condensation particle counters (CPCs) with different dp50 (50% lower size detection efficiency diameter or "cut-off diameter") were conducted on board the M-55 Geophysica in the altitude range of 12.0–20.5 km and on board the DLR Falcon-20 at up to 11.5 km altitude. On board the NASA WB-57F size distributions were measured over Central America in the 4 to 1000 nm diameter range with a system of nucleation mode aerosol spectrometers. Nucleation mode particle concentrations (NNM) were derived from these measurements which allow for identifying many NPF events with NNM in the range of thousands of particles per cm3. Over Australia and West Africa, we identified NPF in the outflow of tropical convection, in particular of a Mesoscale Convective System (MCS). Newly formed particles with NNM > 1000 cm−3 were found to coexist with ice cloud particles (dp > 2 μm) as long as cloud particle concentrations remained below 2 cm−3. The occurrence of NPF within the upper troposphere and the TTL was generally confined within 340 K to 380 K potential temperature, but NPF was of particular strength between 350 K and 370 K (i.e. ~1–4 km below the cold point tropopause). Analyses of the aerosol volatility (at 250 °C) show that in the TTL on average 75–90% of the particles were volatile, compared to typically only 50% in the extra-tropical UT, indicative for the particles to mainly consist of H2SO4-H2O and possibly organic compounds. Along two flight segments over Central and South America (24 February 2005 and 7 August 2006, at 12.5 km altitude) in cloud free air, above thin cirrus, particularly high NNM were observed. Recent lifting had influenced the probed air masses, and NNM reached up to 16 000 particles cm−3 (ambient concentration). A sensitivity study using an aerosol model, which includes neutral and ion induced nucleation processes, simulates NNM in reasonable agreement with the in situ observations of clear-air NPF. Based on new, stringent multi-CPC criteria, our measurements corroborate the hypothesis that the tropical UT and the TTL are regions supplying freshly nucleated particles. Our findings narrow the altitude of the main source region to the bottom TTL, i.e. to the level of main tropical convection outflow, and, by means of measurements of carbon monoxide, they indicate the importance of anthropogenic emissions in NPF. After growth and/or coalescence the nucleation mode particles may act as cloud condensation nuclei in the tropical UT, or, upon ascent into the stratosphere, contribute to maintain the stratospheric background aerosol.

Abstract Background: Patients (pts) with CLL harboring 17p deletion [del(17p)] are considered to have very poor prognosis. Venetoclax (VEN) is an orally bioavailable, selective BCL-2 inhibitor that induces apoptosis in CLL cells independent of p53. A phase 1 study of VEN showed high response rates in pts with relapsed/refractory (R/R) CLL, including del(17p) CLL (overall response rate, ORR = 77%). This pivotal phase 2, single-arm, multicenter study evaluated VEN monotherapy in pts with R/R del(17p) CLL. Methods: Pts with R/R del(17p) CLL, assessed in peripheral blood (PB) by a central laboratory ( >7% cells by Vysis FISH probe), commenced VEN once daily with a weekly dose ramp-up schedule (20, 50, 100, 200, 400 mg) over a period of 5 weeks with tumor lysis syndrome (TLS) prophylaxis. Pts were treated with daily 400 mg continuous dosing until disease progression or discontinuation for another reason. The primary objective was to determine the ORR. Responses were determined by both an independent review committee (IRC) and investigators using iwCLL 2008 criteria. Efficacy analyses were pre-specified to occur once pts completed 36 weeks of VEN, had disease progression, or permanently discontinued. Secondary objectives included CR and PR rates, time to first response, duration of response (DoR), progression-free survival (PFS), overall survival (OS), the proportion of pts proceeding to allogeneic stem cell transplant (allo-SCT), and safety. The level of minimal residual disease (MRD) in PB and/or bone marrow (BM) was assessed in a subset of pts by multi-color flow cytometry using iwCLL-recommended sensitivity criteria of <10-4. Results: A total of 107 pts were enrolled in the main cohort (June 2013–June 2014). Median (range) age was 67 (37–85) years; 65% were male. Median number of prior regimens was 2 (1–10); 78 pts (72.9%) had received prior fludarabine (F), 34 (37.4%) were F-refractory, 54 (50.5%) received prior bendamustine (B), and 27 (50%) were B-refractory. 45 pts (42.1%) were high-risk for TLS based on lymph nodes ≥10 cm (or nodes ≥5 cm with ALC ≥25x109/L). All pts but one had del(17p); 60 of 83 pts with available data (72.3%) had mutated TP53 (investigator reported). As of the data cut-off (April 30, 2015), the median time on study was 12.1 (0.03–21.5) months. The primary endpoint of IRC-assessed ORR was 79.4% (95% CI: 70.5%–86.6%). Deep responses included 7.5% CR/CRi and 2.8% nPR, by IRC (Table). Among pts who achieved PR (69.2%, excluding nPR) or non-responders (20.6%) by IRC, 17 pts (15.9%) had no morphological evidence of CLL in the BM. Investigator-assessed ORR is also reported in the Table. 45 pts had an MRD assessment. Notably 18 pts (17% of whole cohort, 21% of responders) had no detectable MRD in the PB; 10 of these were also tested in BM, 6 were MRD-negative. Median time-to-first response was 0.8 months (0.1–8.1); median time to CR/CRi was 8.2 months (3.0–16.3) by IRC. Overall median DoR, PFS, and OS were not reached. The actuarial 12-month PFS and OS rates were 72.0% and 86.7%, respectively (actuarial 12-month DoR rates in Table). 37 pts discontinued treatment: 22 due to PD (9 Richter's transformation), 9 due to AE, 2 withdrew consent, and 1 with non-compliance; 3 pts proceeded to allo-SCT (2 PR, 1 CR by IRC at time of transplant). 11 deaths occurred (≤30 days from last dose of VEN): 7 due to PD, 4 due to AE (stroke, liver derangement, septic shock, and cardio-respiratory insufficiency). 7 additional deaths occurred beyond 30 days from VEN discontinuation (39-328 days) due to PD. Treatment-emergent AEs (all grades) in ≥20% pts were neutropenia (43%), diarrhea (29%), nausea (29%), anemia (27%), and fatigue (22%). Grade 3/4 AEs in ≥10% pts were neutropenia (40%; 25 pts had grade 4), anemia (18%), and thrombocytopenia (15%). 22.4% of pts had neutropenia (any grade) at study entry. Infection ≥ grade 3 occurred in 20% of pts; most common was pneumonia (5%). Laboratory TLS was reported in 5 pts; none had clinical consequences, and all were manageable with electrolyte management and 1-day dose interruption (2 pts). Conclusions: VEN monotherapy achieved a high ORR and sustained remissions with acceptable toxicity in this ultra-high risk pt population with R/R del(17p) CLL. Undetectable MRD was observed in >20% of responders. More than 10% of all pts achieved independently assessed deep responses (CR, CRi, or nPR). Such depths of response have not been previously reported for this population. Disclosures: Stilgenbauer: AbbVie, Amgen, Boehringer-Ingelheim, Celgene, Genentech, Genzyme, Gilead, GSK, Janssen, Mundipharma, Novartis, Pharmacyclics, Hoffman La Roche: Honoraria , Membership on an entity’s Board of Directors or advisory committees , Research Funding. Off Label Use: Venetoclax is an investigational drug and has no label at this time. Eichhorst:AbbVie, Roche: Membership on an entity’s Board of Directors or advisory committees , Research Funding , Speakers Bureau. Schetelig:GSK, Sanofi, Janssen, Neovii: Membership on an entity’s Board of Directors or advisory committees , Research Funding. Coutre:AbbVie: Research Funding ; Gilead: Research Funding ; Pharmacyclics LLC, an AbbVie Company: Consultancy , Membership on an entity’s Board of Directors or advisory committees , Research Funding ; Janssen: Consultancy , Honoraria , Membership on an entity’s Board of Directors or advisory committees ; Celgene Corporation: Membership on an entity’s Board of Directors or advisory committees , Research Funding ; Celgene Business Advisory Board: Membership on an entity’s Board of Directors or advisory committees ; Novartis: Research Funding. Seymour:Gilead: Honoraria , Membership on an entity’s Board of Directors or advisory committees ; Genentech, Inc.: Membership on an entity’s Board of Directors or advisory committees ; Celgene: Consultancy , Honoraria , Membership on an entity’s Board of Directors or advisory committees , Other: Travel support , Speakers Bureau ; Incyte: Honoraria , Membership on an entity’s Board of Directors or advisory committees ; AbbVie: Consultancy , Honoraria , Membership on an entity’s Board of Directors or advisory committees , Other: Travel support , Research Funding , Speakers Bureau ; Janssen: Honoraria , Membership on an entity’s Board of Directors or advisory committees , Research Funding ; Phebra: Consultancy , Honoraria , Membership on an entity’s Board of Directors or advisory committees ; Takeda: Honoraria , Membership on an entity’s Board of Directors or advisory committees ; Roche: Consultancy , Honoraria , Membership on an entity’s Board of Directors or advisory committees , Other: Travel support , Research Funding ; Infinity: Honoraria , Membership on an entity’s Board of Directors or advisory committees. Puvvada:Genentech, AbbVie, Spectrum, Janssen and Takeda, Pharmacyclics, Genentech/Roche: Consultancy , Membership on an entity’s Board of Directors or advisory committees , Other: Travel funding for Investigators meeting , Research Funding. Wendtner:Mundipharma: Consultancy , Other: travel grants , Research Funding ; Celege: Consultancy , Other: Travel grants , Research Funding ; Gilead: Consultancy , Other: travel grants , Research Funding ; Glaxo-SmithKline: Consultancy , Other: travel grants , Research Funding ; Janssen-Cilag: Consultancy , Other: travel grants , Research Funding ; Pharmacyclics: Consultancy , Other: travel grants , Research Funding ; Hoffmann-LaRoche: Consultancy , Other: travel grants , Research Funding ; Genentech: Consultancy , Other: travel grants , Research Funding ; AbbVie: Consultancy , Other: travel grants , Research Funding. Roberts:AbbVie and Genentech: Research Funding ; Walter and Eliza Hall Institute of Medical Research: Employment. Jurczak:CELLTRION, Inc,: Research Funding ; Celgene, Eisai, Gilead, Janssen, Mundipharma, Pharmacyclics, Pfizer, Roche, Sandoz –Novartis, Spectrum, Takeda, AbbVie, Morphosys, Janssen, Mundipharma, Sandoz –Novartis, Spectrum, Takeda, Teva, Morphosys: Membership on an entity’s Board of Directors or advisory committees , Research Funding. Mulligan:Sanofi Aventis: Research Funding ; Celgene: Consultancy , Honoraria ; Roche: Consultancy , Honoraria , Research Funding , Speakers Bureau ; Janssen: Consultancy , Honoraria , Speakers Bureau ; AbbVie: Membership on an entity’s Board of Directors or advisory committees. Boettcher:AbbVie: Honoraria , Research Funding ; Celgene: Research Funding ; Roche: Honoraria , Other: travel grants , Research Funding. Mobasher:Genentech, Inc.: Employment ; Roche: Equity Ownership. Zhu:AbbVie: Employment , Equity Ownership. Chyla:AbbVie: Employment , Equity Ownership. Verdugo:AbbVie: Employment , Equity Ownership. Enschede:AbbVie: Employment , Equity Ownership. Cerri:AbbVie: Employment , Equity Ownership. Humerickhouse:AbbVie: Employment , Equity Ownership. Gordon:AbbVie: Employment , Equity Ownership. Hallek:GSK, Genentech: Membership on an entity’s Board of Directors or advisory committees , Research Funding ; Celgene: Honoraria , Other: Speakers Bureau and/or Advisory Boards , Research Funding ; Janssen: Honoraria , Other: Speakers Bureau and/or Advisory Boards , Research Funding ; Mundipharma: Honoraria , Other: Speakers Bureau and/or Advisory Boards , Research Funding ; Boehringher Ingelheim: Honoraria , Other: Speakers Bureau and/or Advisory Boards ; Pharmacyclics: Honoraria , Other: Speakers Bureau and/or Advisory Boards , Research Funding ; Roche: Honoraria , Other: Speakers Bureau and/or Advisory Boards , Research Funding ; Gilead: Honoraria , Other: Speakers Bureau and/or Advisory Boards , Research Funding ; AbbVie: Honoraria , Other: Speakers Bureau and/or Advisory Boards , Research Funding. Wierda:AbbVie, Genentech: Consultancy , Research Funding.

Accumulating evidence has confirmed that inherited genetic variations play multi-dimensional roles in childhood acute lymphoblastic leukemia (ALL), i.e., leukemia susceptibility, treatment response, chemotherapy tolerance, and relapse. Germline variants at the GATA3 locus increase the risk of developing Philadelphia chromosome-like ALL (Ph-like ALL) and inferior outcomes in childhood B-ALL among European and American cohorts. However, the role of inherited GATA3 variants in Han Chinese children with ALL still remains unclear. To primarily identify the association of inherited GATA3 variants with treatment response, we retrospectively collected 273 childhood B-ALL blood samples after complete remission was achieved according to the Chinese Childhood Cancer Group ALL 2015. We then genotyped rs3824662 and rs3781093 in the GATA3 locus. The risk allele frequencies of rs3824662 and rs3781093 were 35.7% and 36.3%, respectively, consistent with the 1000 Genomes Project. Using a logistic regression model, we correlated the GATA3 genotype with minimal residual disease (MRD) level. In our single center, we found that GATA3 rs3824662 A allele and rs3781093 C allele statistically associated with positive MRD (the cut-off was &gt;=0.01%, P=0.046 and 0.038, respectively). The A allele in rs3824662 and C allele in rs3781093 linked to 2-fold increase in the risk of MRD compared with their wildtype allele. To explore the biological functions of these two germline SNP variants, we first utilized luciferase reporter assay to determine the impact of GATA3 variants on its transcription activity. Interestingly, the rs3824662 risk A allele significantly increased enhancer activity, while the rs3781093 did not show any effect. We next genetically modified rs3824662 from wild-type C allele to A allele in the lymphoblastoid cell GM12878 using clustered regularly interspaced short palindromic repeats/associated 9 (CRISPR/Cas9) gene editing system. Compared with wildtype GM12878 cells, ~3-folds higher GATA3 transcription level was found in GM12878 cells with A/A or A/C genotype. Integrating the high risk of MRD and upregulated GATA3 expression, we proposed that GATA3 rs3824662 A allele might contribute to poor treatment response by promoting GATA3 transcription. To clarify the association of GATA3 expression with the sensitivity of ALL chemotherapeutic drugs, we retrieved GSE653 and GSE654 expression data for analysis and found that high GATA3 expression significantly correlated with L-asparaginase (L-Asp) and daunorubicin (DNR) resistance. To further confirm the correlation, we ectopically overexpressed GATA3 in B-ALL cell line (Nalm6) and only L-asp resistance was validated. L-asp resistance induced by GATA3 over-expression was rescued by GATA3 interference, consolidating the association between GATA3 and L-asp resistance in B-ALL cells. Next, we probed the mechanism of GATA3-mediated L-asp resistance in B-ALL. We analyzed the association between inherited GATA3 variants and L-asp allergy, and did not identify statistical significance. Meanwhile, we couldn't find the correlation between GATA3 and ASNS, suggesting that ASNS might not be the cause either. Intriguingly, we found GATA3 over-expression induced the activation of autophagy-related genes, BECN1 and ATG5, which was reported to be associated with L-Asp resistance. Tests on primary B-ALL samples further confirmed this findings. To further confirm the effect of GATA3 on autophagy, we cloned highly conserved sequence BECN1 or ATG5 promoter region into luciferase reporter constructs. The Over-expression of GATA3 dramatically increased luciferase activity compared with the corresponding empty vector (P = 0.0098 and 0.0114, respectively), indicating that GATA3 expression were functionally activate their transcription. Taken together, all these data indicated that higher GATA3 expression might induce L-Asp resistance in B-ALL cells via autophagy activation. In conclusion, we first identified that GATA3 rs3824662 associated with the risk of MRD in the childhood ALL cohorts of Han ethnicity. Mechanistic study showed that inherited GATA3 variants possibly contributed to L-Asp resistance via autophagy activation induced by promoting GATA3 enhancer activity, providing new insights into the rationale for the future development of combinational treatment of L-Asp and anti-autophagy regimen in ALL patients. Figure Disclosures No relevant conflicts of interest to declare.

Environments containing significant concentration of NaCl such as salt lakes harbor extremophiles microorganisms which have a great biotechnology interest. To explore the diversity of Bacteria in Chott Tinsilt (Algeria), an isolation program was performed. Water samples were collected from the saltern during the pre-salt harvesting phase. This Chott is high in salt (22.47% (w/v). Seven halophiles Bacteria were selected for further characterization. The isolated strains were able to grow optimally in media with 10–25% (w/v) total salts. Molecular identification of the isolates was performed by sequencing the 16S rRNA gene. It showed that these cultured isolates included members belonging to the Halomonas, Staphylococcus, Salinivibrio, Planococcus and Halobacillus genera with less than 98% of similarity with their closest phylogenetic relative. The halophilic bacterial isolates were also characterized for the production of biosurfactant and industrially important enzymes. Most isolates produced hydrolases and biosurfactants at high salt concentration. In fact, this is the first report on bacterial strains (A4 and B4) which were a good biosurfactant and coagulase producer at 20% and 25% ((w/v)) NaCl. In addition, the biosurfactant produced by the strain B4 at high salinity (25%) was also stable at high temperature (30-100°C) and high alkalinity (pH 11).Key word: Salt Lake, Bacteria, biosurfactant, Chott, halophiles, hydrolases, 16S rRNAINTRODUCTIONSaline lakes cover approximately 10% of the Earth’s surface area. The microbial populations of many hypersaline environments have already been studied in different geographical regions such as Great Salt Lake (USA), Dead Sea (Israel), Wadi Natrun Lake (Egypt), Lake Magadi (Kenya), Soda Lake (Antarctica) and Big Soda Lake and Mono Lake (California). Hypersaline regions differ from each other in terms of geographical location, salt concentration and chemical composition, which determine the nature of inhabitant microorganisms (Gupta et al., 2015). Then low taxonomic diversity is common to all these saline environments (Oren et al., 1993). Halophiles are found in nearly all major microbial clades, including prokaryotic (Bacteria and Archaea) and eukaryotic forms (DasSarma and Arora, 2001). They are classified as slight halophiles when they grow optimally at 0.2–0.85 M (2–5%) NaCl, as moderate halophiles when they grow at 0.85–3.4 M (5–20%) NaCl, and as extreme halophiles when they grow at 3.4–5.1 M (20–30%) NaCl. Hyper saline environments are inhabited by extremely halophilic and halotolerant microorganisms such as Halobacillus sp, Halobacterium sp., Haloarcula sp., Salinibacter ruber , Haloferax sp and Bacillus spp. (Solomon and Viswalingam, 2013). There is a tremendous demand for halophilic bacteria due to their biotechnological importance as sources of halophilic enzymes. Enzymes derived from halophiles are endowed with unique structural features and catalytic power to sustain the metabolic and physiological processes under high salt conditions. Some of these enzymes have been reported to be active and stable under more than one extreme condition (Karan and Khare, 2010). Applications are being considered in a range of industries such as food processing, washing, biosynthetic processes and environmental bioremediation. Halophilic proteases are widely used in the detergent and food industries (DasSarma and Arora, 2001). However, esterases and lipases have also been useful in laundry detergents for the removal of oil stains and are widely used as biocatalysts because of their ability to produce pure compounds. Likewise, amylases are used industrially in the first step of the production of high fructose corn syrup (hydrolysis of corn starch). They are also used in the textile industry in the de-sizing process and added to laundry detergents. Furthermore, for the environmental applications, the use of halophiles for bioremediation and biodegradation of various materials from industrial effluents to soil contaminants and accidental spills are being widely explored. In addition to enzymes, halophilic / halotolerants microorganisms living in saline environments, offer another potential applications in various fields of biotechnology like the production of biosurfactant. Biosurfactants are amphiphilic compounds synthesized from plants and microorganisms. They reduce surface tension and interfacial tension between individual molecules at the surface and interface respectively (Akbari et al., 2018). Comparing to the chemical surfactant, biosurfactant are promising alternative molecules due to their low toxicity, high biodegradability, environmental capability, mild production conditions, lower critical micelle concentration, higher selectivity, availability of resources and ability to function in wide ranges of pH, temperature and salinity (Rocha et al., 1992). They are used in various industries which include pharmaceuticals, petroleum, food, detergents, cosmetics, paints, paper products and water treatment (Akbari et al., 2018). The search for biosurfactants in extremophiles is particularly promising since these biomolecules can adapt and be stable in the harsh environments in which they are to be applied in biotechnology.OBJECTIVESEastern Algeria features numerous ecosystems including hypersaline environments, which are an important source of salt for food. The microbial diversity in Chott Tinsilt, a shallow Salt Lake with more than 200g/L salt concentration and a superficies of 2.154 Ha, has never yet been studied. The purpose of this research was to chemically analyse water samples collected from the Chott, isolate novel extremely or moderate halophilic Bacteria, and examine their phenotypic and phylogenetic characteristics with a view to screening for biosurfactants and enzymes of industrial interest.MATERIALS AND METHODSStudy area: The area is at 5 km of the Commune of Souk-Naâmane and 17 km in the South of the town of Aïn-Melila. This area skirts the trunk road 3 serving Constantine and Batna and the railway Constantine-Biskra. It is part the administrative jurisdiction of the Wilaya of Oum El Bouaghi. The Chott belongs to the wetlands of the High Plains of Constantine with a depth varying rather regularly without never exceeding 0.5 meter. Its length extends on 4 km with a width of 2.5 km (figure 1).Water samples and physico-chemical analysis: In February 2013, water samples were collected from various places at the Chott Tinsilt using Global Positioning System (GPS) coordinates of 35°53’14” N lat. and 06°28’44”E long. Samples were collected randomly in sterile polythene bags and transported immediately to the laboratory for isolation of halophilic microorganisms. All samples were treated within 24 h after collection. Temperature, pH and salinity were measured in situ using a multi-parameter probe (Hanna Instruments, Smithfield, RI, USA). The analytical methods used in this study to measure ions concentration (Ca2+, Mg2+, Fe2+, Na+, K+, Cl−, HCO3−, SO42−) were based on 4500-S-2 F standard methods described elsewhere (Association et al., 1920).Isolation of halophilic bacteria from water sample: The media (M1) used in the present study contain (g/L): 2.0 g of KCl, 100.0/200.0 g of NaCl, 1.0 g of MgSO4.7HO2, 3.0 g of Sodium Citrate, 0.36 g of MnCl2, 10.0 g of yeast extract and 15.0 g agar. The pH was adjusted to 8.0. Different dilutions of water samples were added to the above medium and incubated at 30°C during 2–7 days or more depending on growth. Appearance and growth of halophilic bacteria were monitored regularly. The growth was diluted 10 times and plated on complete medium agar (g/L): glucose 10.0; peptone 5.0; yeast extract 5.0; KH2PO4 5.0; agar 30.0; and NaCl 100.0/200.0. Resultant colonies were purified by repeated streaking on complete media agar. The pure cultures were preserved in 20% glycerol vials and stored at −80°C for long-term preservation.Biochemical characterisation of halophilic bacterial isolates: Bacterial isolates were studied for Gram’s reaction, cell morphology and pigmentation. Enzymatic assays (catalase, oxidase, nitrate reductase and urease), and assays for fermentation of lactose and mannitol were done as described by Smibert (1994).Optimization of growth conditions: Temperature, pH, and salt concentration were optimized for the growth of halophilic bacterial isolates. These growth parameters were studied quantitatively by growing the bacterial isolates in M1 medium with shaking at 200 rpm and measuring the cell density at 600 nm after 8 days of incubation. To study the effect of NaCl on the growth, bacterial isolates were inoculated on M1 medium supplemented with different concentration of NaCl: 1%-35% (w/v). The effect of pH on the growth of halophilic bacterial strains was studied by inoculating isolates on above described growth media containing NaCl and adjusted to acidic pH of 5 and 6 by using 1N HCl and alkaline pH of 8, 9, 10, 11 and 12 using 5N NaOH. The effect of temperature was studied by culturing the bacterial isolates in M1 medium at different temperatures of incubation (4°C–55°C).Screening of halophilic bacteria for hydrolytic enzymes: Hydrolase producing bacteria among the isolates were screened by plate assay on starch, tributyrin, gelatin and DNA agar plates respectively for amylase, lipase, protease and DNAse activities. Amylolytic activity of the cultures was screened on starch nutrient agar plates containing g/L: starch 10.0; peptone 5.0; yeast extract 3.0; agar 30.0; NaCl 100.0/250.0. The pH was 7.0. After incubation at 30 ºC for 7 days, the zone of clearance was determined by flooding the plates with iodine solution. The potential amylase producers were selected based on ratio of zone of clearance diameter to colony diameter. Lipase activity of the cultures was screened on tributyrin nutrient agar plates containing 1% (v/v) of tributyrin. Isolates that showed clear zones of tributyrin hydrolysis were identified as lipase producing bacteria. Proteolytic activity of the isolates was similarly screened on gelatin nutrient agar plates containing 10.0 g/L of gelatin. The isolates showing zones of gelatin clearance upon treatment with acidic mercuric chloride were selected and designated as protease producing bacteria. The presence of DNAse activity on plates was determined on DNAse test agar (BBL) containing 10%-25% (w/v) total salt. After incubation for 7days, the plates were flooded with 1N HCl solution. Clear halos around the colonies indicated DNAse activity (Jeffries et al., 1957).Milk clotting activity (coagulase activity) of the isolates was also determined following the procedure described (Berridge, 1952). Skim milk powder was reconstituted in 10 mM aqueous CaCl2 (pH 6.5) to a final concentration of 0.12 kg/L. Enzyme extracts were added at a rate of 0.1 mL per mL of milk. The coagulation point was determined by manual rotating of the test tube periodically, at short time intervals, and checking for visible clot formation.Screening of halophilic bacteria for biosurfactant production. Oil spread Assay: The Petridis base was filled with 50 mL of distilled water. On the water surface, 20μL of diesel and 10μl of culture were added respectively. The culture was introduced at different spots on the diesel, which is coated on the water surface. The occurrence of a clear zone was an indicator of positive result (Morikawa et al., 2000). The diameter of the oil expelling circles was measured by slide caliber (with a degree of accuracy of 0.02 mm).Surface tension and emulsification index (E24): Isolates were cultivated at 30 °C for 7 days on the enrichment medium containing 10-25% NaCl and diesel oil as the sole carbon source. The medium was centrifuged (7000 rpm for 20 min) and the surface tension of the cell-free culture broth was measured with a TS90000 surface tensiometer (Nima, Coventry, England) as a qualitative indicator of biosurfactant production. The culture broth was collected with a Pasteur pipette to remove the non-emulsified hydrocarbons. The emulsifying capacity was evaluated by an emulsification index (E24). The E24 of culture samples was determined by adding 2 mL of diesel oil to the same amount of culture, mixed for 2 min with a vortex, and allowed to stand for 24 h. E24 index is defined as the percentage of height of emulsified layer (mm) divided by the total height of the liquid column (mm).Biosurfactant stability studies : After growth on diesel oil as sole source of carbone, cultures supernatant obtained after centrifugation at 6,000 rpm for 15 min were considered as the source of crude biosurfactant. Its stability was determined by subjecting the culture supernatant to various temperature ranges (30, 40, 50, 60, 70, 80 and 100 °C) for 30 min then cooled to room temperature. Similarly, the effect of different pH (2–11) on the activity of the biosurfactant was tested. The activity of the biosurfactant was investigated by measuring the emulsification index (El-Sersy, 2012).Molecular identification of potential strains. DNA extraction and PCR amplification of 16S rDNA: Total cellular DNA was extracted from strains and purified as described by Sambrook et al. (1989). DNA was purified using Geneclean® Turbo (Q-BIO gene, Carlsbad, CA, USA) before use as a template in polymerase chain reaction (PCR) amplification. For the 16S rDNA gene sequence, the purified DNA was amplified using a universal primer set, forward primer (27f; 5′-AGA GTT TGA TCM TGG CTC AG) and a reverse primer (1492r; 5′-TAC GGY TAC CTT GTT ACG ACT T) (Lane, 1991). Agarose gel electrophoresis confirmed the amplification product as a 1400-bp DNA fragment.16S rDNA sequencing and Phylogenic analysis: Amplicons generated using primer pair 27f-1492r was sequenced using an automatic sequencer system at Macrogene Company (Seoul, Korea). The sequences were compared with those of the NCBI BLAST GenBank nucleotide sequence databases. Phylogenetic trees were constructed by the neighbor-joining method using MEGA version 5.05 software (Tamura et al., 2011). Bootstrap resembling analysis for 1,000 replicates was performed to estimate the confidence of tree topologies.Nucleotide sequence accession numbers: The nucleotide sequences reported in this work have been deposited in the EMBL Nucleotide Sequence Database. The accession numbers are represented in table 5.Statistics: All experiments were conducted in triplicates. Results were evaluated for statistical significance using ANOVA.RESULTSPhysico-chemical parameters of the collected water samples: The physicochemical properties of the collected water samples are reported in table 1. At the time of sampling, the temperature was 10.6°C and pH 7.89. The salinity of the sample, as determined in situ, was 224.70 g/L (22,47% (w/v)). Chemical analysis of water sample indicated that Na +and Cl- were the most abundant ions (table 1). SO4-2 and Mg+2 was present in much smaller amounts compared to Na +and Cl- concentration. Low levels of calcium, potassium and bicarbonate were also detected, often at less than 1 g/L.Characterization of isolates. Morphological and biochemical characteristic feature of halophilic bacterial isolates: Among 52 strains isolated from water of Chott Tinsilt, seven distinct bacteria (A1, A2, A3, A4, B1, B4 and B5) were chosen for further characterization (table 2). The colour of the isolates varied from beige, pale yellow, yellowish and orange. The bacterial isolates A1, A2, A4, B1 and B5 were rod shaped and gram negative (except B5), whereas A3 and B4 were cocci and gram positive. All strains were oxidase and catalase positive except for B1. Nitrate reductase and urease activities were observed in all the bacterial isolates, except B4. All the bacterial isolates were negative for H2S formation. B5 was the only strain positive for mannitol fermentation (table 2).We isolated halophilic bacteria on growth medium with NaCl supplementation at pH 7 and temperature of 30°C. We studied the effect of NaCl, temperature and pH on the growth of bacterial isolates. All the isolates exhibited growth only in the presence of NaCl indicating that these strains are halophilic. The optimum growth of isolates A3 and B1 was observed in the presence of 10% NaCl, whereas it was 15% NaCl for A1, A2 and B5. A4 and B4 showed optimum growth in the presence of 20% and 25% NaCl respectively. A4, B4 and B5 strains can tolerate up to 35% NaCl.The isolate B1 showed growth in medium supplemented with 10% NaCl and pH range of 7–10. The optimum pH for the growth B1 was 9 and they did not show any detectable growth at or below pH 6 (table 2), which indicates the alkaliphilic nature of B1 isolate. The bacterial isolates A1, A2 and A4 exhibited growth in the range of pH 6–10, while A3 and B4 did not show any growth at pH greater than 8. The optimum pH for growth of all strains (except B1) was pH 7.0 (table 2). These results indicate that A1, A2, A3, A4, B4 and B5 are neutrophilic in nature. All the bacterial isolates exhibited optimal growth at 30°C and no detectable growth at 55°C. Also, detectable growth of isolates A1, A2 and A4 was observed at 4°C. However, none of the bacterial strains could grow below 4°C and above 50°C (table 2).Screening of the halophilic enzymes: To characterize the diversity of halophiles able to produce hydrolytic enzymes among the population of microorganisms inhabiting the hypersaline habitats of East Algeria (Chott Tinsilt), a screening was performed. As described in Materials and Methods, samples were plated on solid media containing 10%-25% (w/v) of total salts and different substrates for the detection of amylase, protease, lipase and DNAse activities. However, coagulase activity was determined in liquid medium using milk as substrate (figure 3). Distributions of hydrolytic activity among the isolates are summarized in table 4.From the seven bacterial isolates, four strains A1, A2, A4 and B5 showed combined hydrolytic activities. They were positive for gelatinase, lipase and coagulase. A3 strain showed gelatinase and lipase activities. DNAse activities were detected with A1, A4, B1 and B5 isolates. B4 presented lipase and coagulase activity. Surprisingly, no amylase activity was detected among all the isolates.Screening for biosurfactant producing isolates: Oil spread assay: The results showed that all the strains could produce notable (>4 cm diameter) oil expelling circles (ranging from 4.11 cm to 4.67 cm). The average diameter for strain B5 was 4.67 cm, significantly (P < 0.05) higher than for the other strains.Surface tension and emulsification index (E24): The assimilation of hydrocarbons as the sole sources of carbon by the isolate strains led to the production of biosurfactants indicated by the emulsification index and the lowering of the surface tension of cell-free supernatant. Based on rapid growth on media containing diesel oil as sole carbon source, the seven isolates were tested for biosurfactant production and emulsification activity. The obtained values of the surface tension measurements as well as the emulsification index (E24) are shown in table 3. The highest reduction of surface tension was achieved with B5 and A3 isolates with values of 25.3 mN m−1 and 28.1 mN m−1 respectively. The emulsifying capacity evaluated by the E24 emulsification index was highest in the culture of isolate B4 (78%), B5 (77%) and A3 (76%) as shown in table 3 and figure 2. These emulsions were stable even after 4 months. The bacteria with emulsification indices higher than 50 % and/or reduction in the surface tension (under 30 mN/m) have been defined as potential biosurfactant producers. Based on surface tension and the E24 index results, isolates B5, B4, A3 and A4 are the best candidates for biosurfactant production. It is important to note that, strains B4 and A4 produce biosurfactant in medium containing respectively 25% and 20% (w/v) NaCl.Stability of biosurfactant activities: The applicability of biosurfactants in several biotechnological fields depends on their stability at different environmental conditions (temperatures, pH and NaCl). For this study, the strain B4 appear very interesting (It can produce biosurfactant at 25 % NaCl) and was choosen for futher analysis for biosurfactant stability. The effects of temperature and pH on the biosurfactant production by the strain B4 are shown in figure 4.biosurfactant in medium containing respectively 25% and 20% (w/v) NaCl.Stability of biosurfactant activities: The applicability of biosurfactants in several biotechnological fields depends on their stability at different environmental conditions (temperatures, pH and NaCl). For this study, the strain B4 appear very interesting (It can produce biosurfactant at 25 % NaCl) and was chosen for further analysis for biosurfactant stability. The effects of temperature and pH on the biosurfactant production by the strain B4 are shown in figure 4. The biosurfactant produced by this strain was shown to be thermostable giving an E-24 Index value greater than 78% (figure 4A). Heating of the biosurfactant to 100 °C caused no significant effect on the biosurfactant performance. Therefore, the surface activity of the crude biosurfactant supernatant remained relatively stable to pH changes between pH 6 and 11. At pH 11, the value of E24 showed almost 76% activity, whereas below pH 6 the activity was decreased up to 40% (figure 4A). The decreases of the emulsification activity by decreasing the pH value from basic to an acidic region; may be due to partial precipitation of the biosurfactant. This result indicated that biosurfactant produced by strain B4 show higher stability at alkaline than in acidic conditions.Molecular identification and phylogenies of potential isolates: To identify halophilic bacterial isolates, the 16S rDNA gene was amplified using gene-specific primers. A PCR product of ≈ 1.3 kb was detected in all the seven isolates. The 16S rDNA amplicons of each bacterial isolate was sequenced on both strands using 27F and 1492R primers. The complete nucleotide sequence of 1336,1374, 1377,1313, 1305,1308 and 1273 bp sequences were obtained from A1, A2, A3, A4, B1, B4 and B5 isolates respectively, and subjected to BLAST analysis. The 16S rDNA sequence analysis showed that the isolated strains belong to the genera Halomonas, Staphylococcus, Salinivibrio, Planococcus and Halobacillus as shown in table 5. The halophilic isolates A2 and A4 showed 97% similarity with the Halomonas variabilis strain GSP3 (accession no. AY505527) and the Halomonas sp. M59 (accession no. AM229319), respectively. As for A1, it showed 96% similarity with the Halomonas venusta strain GSP24 (accession no. AY553074). B1 and B4 showed for their part 96% similarity with the Salinivibrio costicola subsp. alcaliphilus strain 18AG DSM4743 (accession no. NR_042255) and the Planococcus citreus (accession no. JX122551), respectively. The bacterial isolate B5 showed 98% sequence similarity with the Halobacillus trueperi (accession no. HG931926), As for A3, it showed only 95% similarity with the Staphylococcus arlettae (accession no. KR047785). The 16S rDNA nucleotide sequences of all the seven halophilic bacterial strains have been submitted to the NCBI GenBank database under the accession number presented in table 5. The phylogenetic association of the isolates is shown in figure 5.DICUSSIONThe physicochemical properties of the collected water samples indicated that this water was relatively neutral (pH 7.89) similar to the Dead Sea and the Great Salt Lake (USA) and in contrast to the more basic lakes such as Lake Wadi Natrun (Egypt) (pH 11) and El Golea Salt Lake (Algeria) (pH 9). The salinity of the sample was 224.70 g/L (22,47% (w/v). This range of salinity (20-30%) for Chott Tinsilt is comparable to a number of well characterized hypersaline ecosystems including both natural and man-made habitats, such as the Great Salt Lake (USA) and solar salterns of Puerto Rico. Thus, Chott Tinsilt is a hypersaline environment, i.e. environments with salt concentrations well above that of seawater. Chemical analysis of water sample indicated that Na +and Cl- were the most abundant ions, as in most hypersaline ecosystems (with some exceptions such as the Dead Sea). These chemical water characteristics were consistent with the previously reported data in other hypersaline ecosystems (DasSarma and Arora, 2001; Oren, 2002; Hacěne et al., 2004). Among 52 strains isolated from this Chott, seven distinct bacteria (A1, A2, A3, A4, B1, B4 and B5) were chosen for phenotypique, genotypique and phylogenetique characterization.The 16S rDNA sequence analysis showed that the isolated strains belong to the genera Halomonas, Staphylococcus, Salinivibrio, Planococcus and Halobacillus. Genera obtained in the present study are commonly occurring in various saline habitats across the globe. Staphylococci have the ability to grow in a wide range of salt concentrations (Graham and Wilkinson, 1992; Morikawa et al., 2009; Roohi et al., 2014). For example, in Pakistan, Staphylococcus strains were isolated from various salt samples during the study conducted by Roohi et al. (2014) and these results agreed with previous reports. Halomonas, halophilic and/or halotolerant Gram-negative bacteria are typically found in saline environments (Kim et al., 2013). The presence of Planococcus and Halobacillus has been reported in studies about hypersaline lakes; like La Sal del Rey (USA) (Phillips et al., 2012) and Great Salt Lake (Spring et al., 1996), respectively. The Salinivibrio costicola was a representative model for studies on osmoregulatory and other physiological mechanisms of moderately halophilic bacteria (Oren, 2006).However, it is interesting to note that all strains shared less than 98.7% identity (the usual species cut-off proposed by Yarza et al. (2014) with their closest phylogenetic relative, suggesting that they could be considered as new species. Phenotypic, genetic and phylogenetic analyses have been suggested for the complete identification of these strains. Theses bacterial strains were tested for the production of industrially important enzymes (Amylase, protease, lipase, DNAse and coagulase). These isolates are good candidates as sources of novel enzymes with biotechnological potential as they can be used in different industrial processes at high salt concentration (up to 25% NaCl for B4). Prominent amylase, lipase, protease and DNAase activities have been reported from different hypersaline environments across the globe; e.g., Spain (Sánchez‐Porro et al., 2003), Iran (Rohban et al., 2009), Tunisia (Baati et al., 2010) and India (Gupta et al., 2016). However, to the best of our knowledge, the coagulase activity has never been detected in extreme halophilic bacteria. Isolation and characterization of crude enzymes (especially coagulase) to investigate their properties and stability are in progress.The finding of novel enzymes with optimal activities at various ranges of salt concentrations is of great importance. Besides being intrinsically stable and active at high salt concentrations, halophilic and halotolerant enzymes offer great opportunities in biotechnological applications, such as environmental bioremediation (marine, oilfiel) and food processing. The bacterial isolates were also characterized for production of biosurfactants by oil-spread assay, measurement of surface tension and emulsification index (E24). There are few reports on biosurfactant producers in hypersaline environments and in recent years, there has been a greater increase in interest and importance in halophilic bacteria for biomolecules (Donio et al., 2013; Sarafin et al., 2014). Halophiles, which have a unique lipid composition, may have an important role to play as surface-active agents. The archae bacterial ether-linked phytanyl membrane lipid of the extremely halophilic bacteria has been shown to have surfactant properties (Post and Collins, 1982). Yakimov et al. (1995) reported the production of biosurfactant by a halotolerant Bacillus licheniformis strain BAS 50 which was able to produce a lipopeptide surfactant when cultured at salinities up to 13% NaCl. From solar salt, Halomonas sp. BS4 and Kocuria marina BS-15 were found to be able to produce biosurfactant when cultured at salinities of 8% and 10% NaCl respectively (Donio et al., 2013; Sarafin et al., 2014). In the present work, strains B4 and A4 produce biosurfactant in medium containing respectively 25% and 20% NaCl. To our knowledge, this is the first report on biosurfactant production by bacteria under such salt concentration. Biosurfactants have a wide variety of industrial and environmental applications (Akbari et al., 2018) but their applicability depends on their stability at different environmental conditions. The strain B4 which can produce biosurfactant at 25% NaCl showed good stability in alkaline pH and at a temperature range of 30°C-100°C. Due to the enormous utilization of biosurfactant in detergent manufacture the choice of alkaline biosurfactant is researched (Elazzazy et al., 2015). On the other hand, the interesting finding was the thermostability of the produced biosurfactant even after heat treatment (100°C for 30 min) which suggests the use of this biosurfactant in industries where heating is of a paramount importance (Khopade et al., 2012). To date, more attention has been focused on biosurfactant producing bacteria under extreme conditions for industrial and commercial usefulness. In fact, the biosurfactant produce by strain B4 have promising usefulness in pharmaceutical, cosmetics and food industries and for bioremediation in marine environment and Microbial enhanced oil recovery (MEOR) where the salinity, temperature and pH are high.CONCLUSIONThis is the first study on the culturable halophilic bacteria community inhabiting Chott Tinsilt in Eastern Algeria. Different genera of halotolerant bacteria with different phylogeneticaly characteristics have been isolated from this Chott. Culturing of bacteria and their molecular analysis provides an opportunity to have a wide range of cultured microorganisms from extreme habitats like hypersaline environments. Enzymes produced by halophilic bacteria show interesting properties like their ability to remain functional in extreme conditions, such as high temperatures, wide range of pH, and high salt concentrations. These enzymes have great economical potential in industrial, agricultural, chemical, pharmaceutical, and biotechnological applications. Thus, the halophiles isolated from Chott Tinsilt offer an important potential for application in microbial and enzyme biotechnology. In addition, these halo bacterial biosurfactants producers isolated from this Chott will help to develop more valuable eco-friendly products to the pharmacological and food industries and will be usefulness for bioremediation in marine environment and petroleum industry.ACKNOWLEDGMENTSOur thanks to Professor Abdelhamid Zoubir for proofreading the English composition of the present paper.CONFLICT OF INTERESTThe authors declare that they have no conflict of interest.Akbari, S., N. H. Abdurahman, R. M. Yunus, F. Fayaz and O. R. Alara, 2018. Biosurfactants—a new frontier for social and environmental safety: A mini review. Biotechnology research innovation, 2(1): 81-90.Association, A. P. H., A. W. W. Association, W. P. C. 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Abstract Background Protein tyrosine kinase 7 (PTK 7) is a membrane receptor, which can be found in various kinds of cancers. In view of this, detection of PTK 7 in the peripheral circulation would be an effective way for the early diagnosis of cancer. Results In this work, a multi-carbon dots and aptamer-based signal amplification ratiometric fluorescence probe was developed. The fluorescence of the aptamer-modified y-CDs and b-CDs were respectively chosen as the detection signal and interior label. The fluorescence of y-CDs was quenched by Fe3O4 and cDNA (complement to aptamer) compound without PTK 7, but recovered by the addition of PTK 7. Then, the free aptamer was cut by DNase I, which amplified the detection signal. The ratiometric fluorescence sensor for PTK 7 was established with the LOD of 0.016 ng mL−1. Conclusions Summary, a multi-carbon dots and aptamer-based signal amplification ratiometric fluorescence probe was developed for the detection of protein tyrosine kinase 7. The developed probe was applied to PTK 7 detection in MCF-7 cells and human serum with satisfying results, thus indicating that this probe has huge potential in clinical practice.

Abstract Moisture content distributions of Scots pine logs in the green state were measured by a novel multi-step procedure. After sample preparation, the transverse sections of the wood surfaces were scanned by an automated scanning device with a fiber optical probe connected to a Fourier transform near-infrared spectroscope. In the course of the measurement sequences, several issues were addressed, such as surface drying, measurement geometry, ease of automation and interconnected data handling. The near-infrared (NIR) data were first modeled separately for heartwood and sapwood by means of multivariate partial least squares regression. The models for moisture content were evaluated by root mean square error of prediction, the result being 0.8% for heartwood and 10% for sapwood. The two models were then applied to the NIR data collected from sets of disks cut from nine logs. The results of the calculated moisture contents were evaluated by methods of descriptive statistics, and they indicated clear differences and trends in the distribution of moisture content in transverse or longitudinal regions of a log. Additionally, inter-tree variation in moisture content was detected.