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The accumulation of DNA damage and the alteration of the DNA damage response (DDR) are critical features of genetic instability that is presumed to be implicated in BCR/ABL1-mediated blastic transformation of chronic myeloid leukemia (CML). The aim of our study was to analyze underlying mechanisms of genetic instability with regard to DNA damage such as DNA double-strand breaks (DSB), DSB repair and DDR signaling during blastic transformation of CML. Immunofluorescence microscopy of γH2AX was performed for quantification of DSB in peripheral blood mononuclear cells (PBMC) of 8 healthy individuals, 24 chronic phase (CP)-CML patients under current/discontinued tyrosine kinase inhibitor (TKI) treatment (21 patients in deep molecular response (DMR), 3 patients in major molecular response (MMR)), 5 CP-CML patients under current/discontinued TKI treatment with loss of MMR, 3 de novo non-treated CP-CML patients and 2 blast phase (BP)-CML patients. In addition, immunofluorescence microscopy of γH2AX/53BP1 was used for semi-quantification of error-prone DSB repair. Furthermore, immunoblotting of p-ATM, p-ATR, p-CHK1, p-CHK2 and p-TP53 was performed in PBMC of CML patients in comparison to PBMC of healthy individuals. Our analysis revealed an increase in numbers of γH2AX foci in PBMC of CP-CML patients under current/discontinued TKI treatment with loss of MMR (1.8 γH2AX foci per PBMC ± 0.4), in PBMC of de novo non-treated CP-CML patients (2.3 γH2AX foci per PBMC ± 0.7) and in PBMC of BP-CML patients (4.9 γH2AX foci per PBMC ± 0.9) as compared to the number of γH2AX foci in PBMC of healthy individuals (1.0 γH2AX foci per PBMC ± 0.1) and in PBMC of CP-CML patients under current/discontinued TKI treatment in DMR/MMR (1.0 γH2AX foci per PBMC ± 0.1) (Figure 1A and B). Analysis of co-localizing γH2AX/53BP1 foci in PBMC suggested progressive activation of error-prone nonhomologous end-joining repair mechanisms during blastic transformation in CML. Signatures of p-ATM, p-ATR, p-CHK1, p-CHK2 and p-TP53 indicated alterations of the DDR. In summary, our data provide evidence for an accumulation of DNA damage in PBMC of CML patients towards BP-CML patients. We hypothesize that ongoing DSB generation, error-prone DSB repair and DDR alterations might be critical mechanisms of blastic transformation in CML. Figure 1 Analysis of γH2AX foci in freshly isolated peripheral blood mononuclear cells (PBMC) of healthy individuals and chronic myeloid leukemia (CML) patients. (A) Exemplary immunofluorescence microscopic images of γH2AX foci (green, Alexa 488) and cell nuclei (blue, DAPI) in PBMC of a healthy individual (HEALTHY#3), a chronic phase CML patient with a deep molecular response to tyrosine kinase inhibitor (CP-CML DMR#16), a de novo non-treated chronic phase CML patient (CP-CML#1) and a blast phase CML patient (BP-CML#2). (B) γH2AX foci levels in PBMC of healthy individuals and in PBMC of CML patients. Figure 1 Disclosures Saussele: Pfizer: Honoraria; Novartis: Honoraria, Research Funding; Incyte: Honoraria, Research Funding; BMS: Honoraria, Research Funding. Fabarius:Novartis: Research Funding.

Centrosomes play important roles in the maintenance of genetic stability and centrosomal aberrations are a hallmark of cancer. Deregulation of centriole duplication leads to supernumerary centrosomes, sister chromatid missegregation and could result in chromosomal instability (CIN) and aneuploidy. CIN is a common feature in at least 45 % of patients with myelodysplastic syndromes (MDS). Therefore, we sought to investigate the centrosomal status and its role for development of CIN in bone marrow (BM) cells of MDS patients. BM cells of 34 MDS patients were examined cytogenetically. Furthermore, cells were immunostained with a centrosome-specific antibody to pericentrin to analyze the centrosomal status. Umbilical cord blood specimens and BM cells of healthy persons (n = 11 and n = 4) served as controls. In addition, the protein expression of the protease separase responsible for genetic stability was examined by western blot analysis. Centrosome abnormalities were detected in 10 % (range, 4–17 %) of cells of MDS samples, but in only 2 % (range, 0–4 %) of cells of healthy controls. Normal karyotypes were found in control cells and in BM cells of 16/34 MDS patients. The incidence of centrosomal alterations was higher in BM cells of patients with cytogenetic alterations (mean, 12 %) compared to BM cells of patients without cytogenetic changes (mean, 7 %). Our results indicate that centrosome alterations are a common and early detectable feature in MDS patients and may contribute to the acquisition of chromosomal aberrations. We assume that centrosome defects could be involved in disease progression and may serve as a future prognostic marker.

Increased DNA damage and alteration of the DNA damage response (DDR) are critical features of genetic instability presumably implicated in pathogenesis of myelodysplastic syndromes (MDS) and acute myeloid leukemias (AML). We used immunofluorescence staining of γH2AX and 53BP1 for analyzing DNA double-strand breaks (DSB) in MDS and AML cell lines, in CD34+ selected cells of normal and MDS bone marrow (including three cases of chronic myelomonocytic leukemias) and in blasts of AML bone marrow. In addition, we screened for activation of the DDR by immunoblotting of p-ATM, p-ATR, p-CHK1, p-CHK2 and p-TP53. As compared to γH2AX foci levels in normal bone marrow samples (0.2 focus per CD34+ cell±0.0; mean±standard error of mean), increased levels of γH2AX foci were detected in 16/16 MDS bone marrow samples (2.8 foci per CD34+ cell±0.5), 18/18 AML bone marrow samples (5.5 foci per blast±0.5), 1/1 MDS cell line (6.4 foci per cell) and 6/6 AML cell lines (12.0 foci per cell±0.6). γH2AX and 53BP1 co-localized in all tested samples forming diffuse, clustered and marginal patterns. Further, DDR proteins were expressed heterogeneously suggesting impairment of the DDR. In summary, our results provide evidence for a continuous increase of DSB across the spectrum from MDS to AML in conjunction with an impaired DDR. Co-localization of γH2AX and 53BP1 indicates promotion of (in)effective nonhomologous end-joining repair mechanisms at sites of DSB. Moreover, γH2AX/53BP1 foci distribution presumably reveals a non-random spatial organization of the genome in MDS and AML.

The aim of this study was to systematically evaluate the cumulative radiation exposure and the associated lifetime-cancer-risk from diagnostic imaging in patients with Hodgkin-lymphoma-(HL) or diffuse-large-B-cell-lymphoma (DLBCL). 99 consecutive patients (53-males) diagnosed with HL or DLBCL were included in the study and followed. Based on the imaging reports, organ and effective-doses-(ED) were calculated individually for each patient and the excess lifetime risks were estimated. The average ED in the first year after diagnosis was significantly different for men (59 ± 33 mSv) and women (744 ± 33 mSv)-(p 0.05). Over all years, more than 90% of the ED resulted from CT. The average cumulative radiation risk estimated for the first year was significantly lower for men (0.76 ± 0.41%) as compared to women (1.28 ± 0.54%)-(p

Multiple electrical incidents occurred during construction and commissioning activities at a new production facility. This paper describes the investigations and findings while highlighting similarities between the incidents, designing for safety, and management of electrical safety during the transfer of construction to commissioning activities.

High mortality rates of invasive fungal disease (IFD), especially invasive aspergillosis (IA), in immunocompromised haematological patients and current diagnostic limitations require improvement of detection of fungal pathogens by defining the optimal use of biomarkers and clinical samples. Concurrent bronchoalveolar lavage (BAL) and peripheral blood samples of 99 haematological patients with suspected IFD were investigated within a multicentre prospective study. Diagnostic performance of a galactomannan (GM) enzyme immune assay (EIA), a 1,3-β-D-glucan assay (BDG), an Aspergillus PCR, and a multifungal DNA-microarray (Chip) alone or in combination were calculated. IFD were classified as proven (n=3), probable (n=34), possible (n=33), and no IFD (n=29) according to EORTC/MSG criteria. GM, PCR, and Chip showed superior diagnostic performance in BAL than in blood, whereas specificity of BDG in BAL was poor (48% (14/29)). The combination of GM (BAL) with BDG (blood) showed sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and DOR (diagnostic odds ratio) of 92% (34/37), 93% (27/29), 94%, 90%, and 153.0, respectively. Combining GM (BAL) with PCR (BAL) showed convincing diagnostic potential for diagnosing IA with sensitivity, specificity, PPV, NPV, and DOR of 85% (17/20), 97% (28/29), 94%, 90%, and 158.7. Addition of the DNA-microarray resulted in further detection of two mucormycetes infections. In 1 out of 15 Aspergillus DNA-positive samples a triazole resistance-mediating Cyp51A mutation was found. Combination of biomarkers is superior to their sole use in diagnosing IFD, particularly IA. Integrating blood and BAL samples into a diagnostic algorithm is an advantageous approach.

Genetic lesions are crucial for cancer initiation. Recently, whole genome sequencing, using next generation technology, was used as a systematic approach to identify mutations in genomes of various types of tumors including melanoma, lung and breast cancer, as well as acute myeloid leukemia (AML). Here, we identify tumor-specific somatic mutations by sequencing transcriptionally active genes. Mutations were detected by comparing the transcriptome sequence of an AML sample with the corresponding remission sample. Using this approach, we found five non-synonymous mutations specific to the tumor sample. They include a nonsense mutation affecting the RUNX1 gene, which is a known mutational target in AML, and a missense mutation in the putative tumor suppressor gene TLE4, which encodes a RUNX1 interacting protein. Another missense mutation was identified in SHKBP1, which acts downstream of FLT3, a receptor tyrosine kinase mutated in about 30% of AML cases. The frequency of mutations in TLE4 and SHKBP1 in 95 cytogenetically normal AML patients was 2%. Our study demonstrates that whole transcriptome sequencing leads to the rapid detection of recurring point mutations in the coding regions of genes relevant to malignant transformation.

Die psychogene Polydipsie mit der Folge einer ausgepragten isovolamischen Hyponatriamie wurde in der Literatur bereits mehrfach beschrieben [4, 8]. Als eine Folge dieser Elektrolytstorung kann es zu einem Hirnodem mit sekundarer Enzephalopathie bis hin zu epileptischen Anfallen kommen. Eine seltene weitere Folge stellt die Rhabdomyolyse dar mit all ihren Komplikationen (Niereninsuffizienz, Hyperkaliamie, Herzrhythmusstorungen) bis hin zum Kompartmentsyndrom bei ausgepragtem Muskelodem. Wir berichten uber einen Patienten mit einer durch eine psychogene Polydipsie induzierte Hyponatriamie, die zu einer Rhabdomyolyse mit daraus resultierendem Kompartmentsyndrom fuhrte.

Internal fixation of displaced fractures of the greater tuberosity allowing functional aftercare.Displaced fractures of the greater tuberosity5 mm. Displaced fractures of the greater tuberosity3 mm in athletes or overhead workers. Multiply fragmented fractures of the greater tuberosity.Displaced 3‑ or 4‑part fractures of the proximal humerus. Nondisplaced fractures of the greater tuberosity.Exposure of the fracture of the greater tuberosity by an anterolateral approach. Open reduction and temporary retention with a Kirschner wire or a "Kugelspieß" or reinforcement of the supraspinatus tendon and distal retention. Bending and positioning of the Bamberg plate and fixation by conventional or locking screws. Optional fixation of the rotator cuff to the plate. Exact monitoring of the implant position using the image intensifier to avoid inadequate distalization of the greater tuberosity.Arm sling (e. g. Gilchrist) for 2 weeks. Start passive assisted exercise on postoperative day 1. Movement allowed up to the pain threshold. Physiotherapeutic treatment to prevent adhesions and capsular shrinking.In all, 10 patients with displaced fractures of the greater tuberosity underwent osteosynthesis using the Bamberg plate. After a follow-up of at least 6 months, a Constant-Murley score of 94.2 points (range 91-98 points) was achieved. The patients' average age was 45.6 years (range 29-68 years).

High concentrations (21–44 mg kg-1 dry matter) of Mo have been identified in forage in several reclaimed mining areas in British Columbia. Since Mo concentrations greater than 5 mg kg-1 in forage dry matter may result in molybdenosis because of a secondary Cu deficiency in ruminants, a study was undertaken to determine if cattle can safely graze the reclaimed land. For 12-wk grazing periods in 1994, 1995 and 1996, 32 cow/calf pairs grazed high-Mo forage at a reclaimed mine tailings site located at the Highland Valley Copper mine near Logan lake, BC. Half of the animals in the trial received a Cu supplement (All-Trace copper bolus) and the other half served as a control group. There were no significant differences (P < 0.05) in weight gain, liver Mo, serum Cu and Mo, and milk Cu and Mo between the two treatment groups of cows. Liver Cu was higher for the Cu bolus group at certain time periods in 1994 and 1995, indicating that the bolus was effective at supplying Cu. At all times, the liver Cu levels for the animals remained above the recommended critical level of 25 mg kg-1 dry matter. Animals appeared healthy and no signs of Cu deficiency were observed. Key words: Molybdenum, molybdenosis, copper, mine tailings, reclamation, animal health

Psychogenic polydipsia leading to severe hyponatremia is well documented in the literature. This electrolyte disorder can result in encephalopathy, cerebral edema and epileptic seizures. Another rare effect is rhabdomyolysis with all its well known complications (e.g. renal failure, hyperkalemia and cardiac arrhythmia) and even resulting in compartment syndrome due to severe muscle edema. We present the case of a patient with severe hyponatremia caused by psychogenic polydipsia leading to rhabdomyolysis and compartment syndrome.

Tyrosine kinase 2 (TYK2) has a pivotal role in immunity to infection and tumor surveillance. It is associated with several cytokine receptor chains including type I interferon (IFN) receptor 1 (IFNAR1), interleukin- (IL-) 12 receptor beta 1 (IL-12Rb1) and IL-10R2. We have generated a mouse with a conditional Tyk2 null allele and proved integrity of the conditional Tyk2 locus. TYK2 was successfully removed by the use of ubiquitous and tissue-specific Cre-expressing mouse strains. Myeloid TYK2 was found to critically contribute to the defense against murine cytomegalovirus. Ubiquitous TYK2 ablation severely impaired tumor immunosurveillance, while deletion in myeloid, dendritic or T cells alone showed no effect. The conditional Tyk2 mouse strain will be instrumental to further dissect TYK2 functions in infection, inflammation and cancer.

A 2 x 3 factorial experiment was conducted to determine the effects of an implant (trenbolene acetate/estradiol or no implant) and method of estrus suppression (immunization against GnRH, melengestrol acetate, or no suppression) on growth performance and carcass characteristics of heifers fed for slaughter. At the start of a 21-d feed adaption phase, crossbred beef heifers (n = 144, 390+/-2.8 kg) were given their first dose of an anti-GnRH vaccine or started on melengestrol acetate (MGA). Thereafter, heifers were fed a high-concentrate diet (78% barley grain) for 84 d (Days 0 to 83), received implants on Day 0, a second vaccination on Day 21, and were slaughtered on Days 84 or 85. Implanting increased average daily gain (1.72 vs 1.50 kg/d, P0.01), feed efficiency (6.02 vs 6.75 kg dry matter intake/kg gain, P0.01), preslaughter weight (532 vs 513 kg, P0.01), carcass weight (301 vs 289 kg, P0.01), and ribeye area (88.6 vs 85.9 cm2, P0.05), but had no affect (P0.05) on dry matter intake, grade fat thickness, marbling score, or lean yield. Compared to heifers fed MGA, those immunized against GnRH had a greater ribeye area (90.0 vs 84.6 cm2) and lean yield (63 vs 61%), and had thinner grade fat (7.5 vs 8.6 mm; P0.05 for each). Furthermore, immunized heifers had lower (P0.001) plasma progesterone concentrations than control heifers on Days 42, 63 and 83. Heifers fed MGA had less estrus mounting activity (P0.05) and lower plasma progesterone concentrations (P0.001) than the remaining heifers. Method of estrus suppression did not affect (P0.05) preslaughter weight, average daily gain, dry matter intake, feed efficiency, carcass weight, or marbling score. In conclusion, implanting significantly increased growth performance and preslaughter and carcass weights. Compared to heifers fed MGA, immunization against GnRH significantly increased ribeye area and lean yield, and reduced grade fat thickness

Condensed tannins (CT) are polyphenolic secondary plant products that are widespread in the plant kingdom. By definition, CT bind to protein and are regarded as "antinutritional" compounds which reduce protein digestibility. Variations in CT chemistry alter protein binding capacities among polymers from different plant species and developmental stages. Condensed tannins interact with proteins in feed, saliva and microbial cells, with microbial exoenzymes, and with endogenous proteins or other feed components, which alters digestive processes as compared with diets free of CT. Tannin levels exceeding 40 to 50 g kg−1 DM in forages may reduce protein and DM digestibility of the forages by ruminants. At low to moderate levels, CT increase the quantity of dietary protein, especially essential amino acids, flowing to the small intestine. Unlike alfalfa, legumes that contain CT do not cause bloat. Dietary CT may provide a means to beneficially manipulate protein digestion and/or prevent pasture bloat in ruminants. Research efforts are also currently being directed to genetically modify alfalfa to de-repress its CT biosynthetic pathway, or to move genes encoding steps of this pathway into alfalfa from other legumes. The effects of CT in forage legumes on digestion in ruminants and the potential use of tannin-rich forages for preventing bloat in grazing systems are discussed. Key words: Tannins, bloat, by-pass protein, cattle, grazing, alfalfa

Alfalfa has been recognized for its superior yield and quality in seeded pastures. However, when grazing immature alfalfa there is a risk of animal losses due to frothy bloat in some ruminant livestock. Inclusion of at least 50% grass in the pasture mixture is commonly recommended to reduce the risk of bloat. Two decades of plant breeding have resulted in the release of AC Grazeland, an alfalfa cultivar that reduces the incidence of bloat. Other bloat control agents such as pluronic detergents and ionophores can also be of value. Development of grazing-tolerant alfalfa varieties is solving some of the problems associated with lack of persistence of alfalfa in mixed stands; however, they are not bloat-safe. Animal productivity commonly increases when alfalfa is included in pasture mixtures. Improvements in cattle rate of gain are observed when alfalfa contributes as little as 35% to the sward. Grazing management is the principal method for controlling pasture yield and quality as well as animal performance and bloat incidence. When grazing management is used to optimize pasture production and nutrient intake, yearling steers can gain as much as 1.5 kg head−1 d−1 and liveweight production ranging from 107 kg ha−1 (on dryland) to 1946 kg ha−1 (under irrigation) can be expected. Limiting utilization of alfalfa-based pasture to ≤70% may be more important for maximizing gain per head than managing herbage quality. Key words: Alfalfa, beef production, forage, grazing

Yearling cattle fed pasture-only for the entire grazing season were compared with cattle receiving pasture plus steam-rolled barley (SRB; 4 kg head−1 d−1) or pasture plus SRB (4 kg head−1 d−1) treated with Tween 80 at a rate of 4 kg t−1, for 74 d prior to slaughter. Heifers were more likely (P = 0.015) to grade A or higher than steers. Feeding supplemental grain on pasture increased pasture gains (P 0.05) improve carcass quality, chemical analysis, shear force, carcass value or gross return per head. Treatment of SRB with Tween-80 did not influence (P > 0.05) cattle performance or carcass quality. From the results reported in this study, it is possible to finish cattle on pasture to Canada A grade standards. Key words: Forage-finishing, carcass quality, beef, pasture, grain

Esophageal fistula and hand-clipped samples were collected from a 32-yr-old crested wheatgrass (Agropyron cristatum L.) pasture and an alfalfa (Medicago sativa L.)/meadow bromegrass (Bromus biebersteinii Roem & Schult.)/Russian wildrye (Psathyrostachys juncea [Fisch.] Nevski) pasture to compare estimates of nutritional quality. Samples were analyzed for crude protein (CP), in vitro digestible organic matter (IVDOM) and selected minerals. Fistula samples had greater (P 0.05) on two occasions and IVDOM did not differ (P > 0.05) on one occasion. Differences in mineral content between clipped and esophageal samples were less consistent, but when they occurred the mineral contents of esophageal samples usually exceeded (P

Barley silage was sprayed with water or with a 2:1 combination of commercial cellulase and xylanase preparations, or the enzymes were introduced directly into the rumen, in a digestibility study (replicated incomplete 3 × 3 Latin square) using 10 sheep. Apparent digestibilities of dry matter (DM) and neutral detergent fibre (NDF) were lower (P 0.05) intake of DM, organic matter or digestible organic matter, or digestibilities of DM or NDF, ruminal pH, xylanase activity, endoglucanase activity or ruminal cellulolytic bacterial populations. Treating the silage portion of an 82.5% barley silage backgrounding diet with the enzyme mix at 0, 1.25, 3.5 or 5.0 L t−1 DM tended to linearly increase (P = 0.08) final weights of steers (n = 24). Average daily gain tended to be (P = 0.06) and feed intake and feed efficiency were (P = 0.04 and P = 0.03, respectively) quadratically related to these enzyme concentrations from days 0 to 56, but not overall (days 0 to 120). In contrast, treatment of both portions (forage and concentrate) of a 70% barley-ryegrass silage finishing diet at 3.5 L t−1 DM increased (P 0.1) by enzyme supplementation. In this study, treating the total mixed ration improved feedlot cattle performance more than treating the silage component alone. Key words: Enzymes, beef cattle, growth, barley silage

The effects of sainfoin (Onobrychis viciifolia) on digestion of alfalfa (Medicago sativa) were investigated in vitro and in vivo. Fresh alfalfa and sainfoin were incubated in an artificial rumen (Rusitec) in ratios of 100:0, 75:25, 50:50, 25:75 and 0:100 (as-fed). Disappearances of dry matter and N from sainfoin were 77 and 65% of those from alfalfa, respectively. Protease and endoglucanase activities, NH3-N and methane production declined (P 0.05), but cells incorporated more 15NH3N as sainfoin in the diet increased. Chopped leaves (100:0, 95:5 and 90:10 alfalfa:sainfoin) were incubated for 48 h with diluted ruminal fluid containing 0 or 50 mg polyethylene glycol, which binds tannins. Gas and volatile fatty acid productions were similar (P > 0.05) across treatments, but including 10% sainfoin (without polyethylene glycol) reduced (P 3 concentrations between 8 and 24 h. Sainfoin tannins reduced degradation of forage protein without affecting the digestibility of the nonprotein fraction. Alfalfa herbage was fed alone or with early- to full-bloom sainfoin herbage (at 10 or 20% of ad libitum alfalfa dry matter intake) or with sainfoin hay or pellets, to eight Jersey steers in crossover trials conducted over 4 yr. Including sainfoin in the diet reduced (P

A number of observations including clinical manifestation, course, outcome, and family history, support the view that patients presenting with a major depression occurring first in late life should be treated as a nosological subgroup. In this study quantitative magnetic resonance imaging (MRI) was used to investigate volumes of different brain structures in 19 patients with late onset major depression (age of onset > 50) and 13 age matched controls. 3-D MRI sequences were acquired using a Siemens 1.5T scanner. Whole brain volume, CSF volume, volume of the frontal and temporal lobes and the volume of the amygdala-hippocampus complex were assessed using the software NMRWin. Compared to the controls, depressed patients showed a significantly lower whole brain volume and a significantly higher CSF volume, whereas volumes of the frontal and temporal lobes as well as the amygdala-hippocampus complex volumes were not significantly decreased. In addition, depressed patients exhibited a higher ventricle-brain ratio suggesting a higher degree of central atrophy compared to healthy individuals. Our results indicate that cerebral changes involving subcortical structures are of relevance in the pathogenesis of late-onset depression. Defining the aetiology of these lesions may be important for the development of preventive treatment of depression in the elderly.

A 4-yr experiment was conducted (1991 to 1994) near Brandon, MB, to determine the effects of grazing system (continuous and rotational) and stocking rate [light (1.1 steers ha−1); heavy (2.2 steers ha−1)] on the productivity, botanical composition and soil surface characteristics of an alfalfa (Medicago sativa L.; approximately 70%), meadow bromegrass (Bromus biebersteinii Roem & Schult.; 25%) and Russian wild ryegrass [Psathyrostachys juncea (Fisch.) Nevski; 5%] pasture. Grazing season length was shorter (P –1) was greater (P −1) and during the season (kg hd−1) at light than at heavy stocking rates, while total liveweight production (kg ha−1) was greater (P 0.05) within grazing systems and stocking rates from 1991 to 1993, but in 1994, production and disappearance were greater (P 0.05) within either grazing system or stocking rate, except in 1992, when basal cover was lowest (P

An experiment was conducted to determine the effects of grazing system (continuous and rotational), stocking rate (light, 1.1 steers ha−1; heavy, 2.2 steers ha−1) and season of use on forage intake and grazing behaviour of stocker cattle grazing an approximately 70% alfalfa (Medicago sativa L.), 25% meadow bromegrass (Bromus biebersteinii Roem & Schult.) and 5% Russian wildrye (Psathyrostachys juncea (Fisch.) Nevski) pasture. To determine organic matter intake, grazed herbage was collected with esophageal fistulated cattle and analyzed for in vitro digestible organic matter while fecal output was determined using chromic oxide. These variables were used to calculate organic matter intake. Daily herbage consumption (g OM kg BW−0.75 d−1 and kg OM d−1) did not differ (P > 0.05) for either grazing system or stocking rate from 1991 to 1993, with the exception of greater (P −1) late in the 1991 season. Grazing time was usually lower (8.9 vs. 10.3 h (3-yr mean); P −1, regardless of year, season, grazing system or stocking rate. Rate of intake (g OM kg−1BW−0.75 h−1) did not differ (P > 0.05) for either grazing system, although it tended to be greater (P

A series of cells representing normal, non-tumorigenic cell lines, as well as differentiating neoplastic and undifferentiated neoplastic rat tracheal epithelial cell populations were evaluated for their ability to establish homologous and/ or heterologous cell-cell gap junction communication in culture. Gap junction communication was evaluated by flow cytometric quantitation of the transfer of the fluorescent dye calcein from a donor to a recipient cell population via gap junctions. The data indicate that normal primary cultures of rat tracheal epithelial cells, as well as non-tumorigenic cell lines and squamous cell carcinomas cell populations, retain the ability to establish both homologous and heterologous gap junction communication. In all cases an average of >48% of recipient cells had acquired calcein label during a 5-h interval of co-culture of donor and recipient cells at confluent densities. Cells harvested directly from squamous cell carcinoma tumors exhibited similar levels of cell-cell communication. In contrast, cells giving rise to undifferentiated carcinomas, as well as cells harvested from undifferentiated carcinomas, exhibited very low levels or no homologous or heterologous cell-cell communication. Cell populations exhibiting distinctly different communication phenotypes were evaluated by Northern blot analysis for expression of connexins (Cx 26, 32 and 43) and E-cadherin. Neither communicating nor non-communicating cells expressed connexin 32. Those cell populations, which established functional gap junctions, expressed E-cadherin as well as connexin 26 and/or 43. In contrast, those cell populations that lacked the ability to communicate universally lacked expression of E-cadherin, and a quarter also lacked expression of detectable levels of connexin.

The effects of estrus suppression (none, oral melengestrol acetate (0.4 mg d−1) or ovariectomy via Willis spay technique) and an anabolic implant (none or Synovex Plus®) on growth performance and carcass characteristics were determined using 90 beef heifers of composite breeding in a 2 × 3 factorial experiment conducted in the last 50 d before slaughter. Spaying caused no morbidity or mortality and resulted in average daily gains (1.37 kg d−1) comparable to heifers fed melengestrol acetate (1.33 kg d−1). Implanting improved (P 0.05) carcass traits. Spaying and implanting represent an effective means of improving returns from heifers, even late in the finishing period. Key words: Ovariectomy, implants, beef heifers, carcass composition

A 3-yr field experiment was conducted (1991–1993) near Brandon, MB to determine the effects of continuous and rotational stocking, both at heavy (2.2 steers ha−1) and light (1.1 steers ha−1) stocking rates, and of season of use on nutritive value and sward characteristics of an alfalfa-grass pasture [approximately 70% alfalfa (Medicago sativa L.), 25% meadow bromegrass (Bromus biebersteinii Roem & Schult.) and 5% Russian wildrye (Psathyrostachys juncea (Fisch.) Nevski)]. Grazed herbage quality and herbage mass were determined concurrently on continuous and rotational stocking treatments several times each year, as animals entered and exited the 5th of 10 rotationally stocked paddocks. Grazing system and stocking rate did not affect (P > 0.05) dietary CP and IVDOM, except once (P 0.05) between grazing systems. As the 1991 season advanced, there was a decline (P 0.05). In 1992, seasonal declines (P 0.05) by grazing system. Significant relationships (P

In this paper, we rotate an array of asymmetrical double layer of 4-strips windmill structure to investigate its effect on the chirality and sensitivity detection of biomolecular structures. The structure is made up of silver material with 300nm pitch and 10nm separation between layers. The spectrum shows two resonance modes on 600THz and 900THz with linear polarized light normally incident on the structure. We investigate the CD by rotating one of the layers with respect to the horizontal axis of the other layer by the angle θ. It is observed that the CD spectra at different angles are different. The rotation resulted in larger wavelength shift of the CD spectra. In addition, the CD also increases with the rotating angle given a larger absorption difference between the left and right handed circular polarized light.

In this paper, the effect of tapered asymmetrical gammadion on the optical rotational properties and sensitivity detection of biomolecular structures is presented. The asymmetrical structure is made up of gold material on a glass structure and immersed in water. The chirality of the un-rotated array is first determined by measuring the circular dichroism (CD) spectrum. Three modes, arising from Bloch periodic theory and surface resonance mode are observed. Then each asymmetrical gammadion structure is tapered at the arms. Tapering fraction, which define the ratio of tapered end to the untapered end is used for defining the new design. The designs are then fabricated using e-beam lithography and tested using the polarimeter for CD spectra. The spectra show that the three CD modes changes in degree amplitude and wavelength, especially for smaller tapering fractions.

The objective of this study was to determine the relative usefulness of physical examination, plain radiographs, and magnetic resonance imaging (MRI) using T1- and T2-weighted spin-echo images in evaluating the extent of urate deposition and soft tissue destruction in gouty arthritis. Seven patients with chronic tophaceous gout of the hands and wrists were examined to identify all clinically apparent tophi. Plain radiographs of the hands and wrist were obtained to further quantify soft tissue and osseous changes. MRI was then performed of the involved areas and a comparison made between soft tissue and bony changes observed by clinical examination and plain radiographs and those observed by MRI. Plain radiographs and physical examination markedly underestimate the size and extent of soft tissue and osseous involvement by tophi when compared with the findings of MRI. MRI also detects early, subclinical tophaceous deposits and indicates that urate deposits appear to spread along compartmental and fascial planes as opposed to the traditional view of strict radial growth. MRI is a useful method of determining the extent of disease in tophaceous gout and may provide information regarding the patterns of deposition and spread of monosodium urate crystals.

Systemic mastocytosis (SM) usually arises as a consequence of an acquired mutation in the receptor tyrosine kinase KIT, usually KIT D816V (>80-90% of patients). The presence of additional molecular aberrations, e.g. in SRSF2, ASXL, or RUNX1 (S/A/Rpos), has a strong adverse impact on disease phenotype and prognosis in patients with advanced SM (advSM) (Jawhar et al., Leukemia 30, 2016). However, little is known about the frequency and prognostic impact of cytogenetic aberrations and potential interactions between cytogenetic and molecular aberrations in the context of prognostication. We therefore analyzed clinical, morphological, cytogenetic and molecular characteristics of 109 patients (KIT D816V, n=102, 94%; KIT D816H, n=2, 2%; no KIT mutation, n=5, 5%) with indolent SM (ISM, n=27) and advSM (n=82) with or without an associated hematologic neoplasm (AHN) [SM-AHN, n=60; aggressive SM, n=3; mast cell leukemia (MCL), n=9; MCL-AHN, n=10)]. Next-Generation Deep Amplicon Sequencing (NGS) was performed to investigate 18 candidate genes as previously described (Schwaab et al., Blood 122, 2013). All ISM patients had a normal karyotype. In advSM, an aberrant karyotype was identified in 16/82 (18%) patients. By analogy with other myeloid neoplasms, e.g. MDS or AML, patients were classified according to their karyotype into two groups. The good-risk group (n=73) included patients with a normal karyotype (n=66) plus those with a favorable karyotype (n=7; del(5q), n=4; trisomy 8, n=1; del(1q), n=1; del(12p), n=1), while the poor-risk group (n=9) included patients with a complex karyotype (defined as ≥3 abnormalities; n=8) or monosomy 7 (n=1). Seven of 9 (78%) patients with a poor-risk karyotype and 2/7 (29%) patients with a favorable karyotype progressed to secondary AML (n=7) or MCL (n=2) within a median observation time of 13 months (range, 0-26). Overall, the median overall survival (OS) of patients with a poor-risk karyotype was significantly shorter than in patients with a good-risk karyotype (4 vs. 41 months; hazard ratio, HR, 9.6, 95% CI 3.9-23.2; P Figure Kaplan-Meier estimates of overall survival (OS) in patients with advanced systemic mastocytosis depending on the karyotype and the mutation status in the SRSF2/ASXL1/RUNX1 gene panel (S/A/R). Pairwise significantly different OS probabilities were observed for the comparison good-risk karyotype + S/A/Rneg (n=27) vs. good-risk karyotype + S/A/Rpos (n=31) vs. poor-risk karyotype (n=9). Figure. Kaplan-Meier estimates of overall survival (OS) in patients with advanced systemic mastocytosis depending on the karyotype and the mutation status in the SRSF2/ASXL1/RUNX1 gene panel (S/A/R). Pairwise significantly different OS probabilities were observed for the comparison good-risk karyotype + S/A/Rneg (n=27) vs. good-risk karyotype + S/A/Rpos (n=31) vs. poor-risk karyotype (n=9). Disclosures Valent: Novartis: Honoraria, Research Funding; Ariad: Honoraria, Research Funding; Amgen: Honoraria; Celgene: Honoraria, Research Funding; Deciphera Pharmaceuticals: Research Funding. Haferlach:MLL Munich Leukemia Laboratory: Employment, Equity Ownership. Meggendorfer:MLL Munich Leukemia Laboratory: Employment.

Purpose: Increased DNA damage and alteration of the DNA damage response (DDR) are critical features of genetic instability presumably implicated in pathogenesis of myelodysplastic syndromes (MDS) and acute myeloid leukemias (AML). Materials and methods: We performed combined γH2AX/53BP1 immunofluorescent focus staining of DNA double-strand breaks (DSB) in MDS and AML cell lines, in CD34+ selected cells of normal and MDS bone marrow (including three cases of chronic myelomonocytic leukemias) and in blasts of AML bone marrow. In addition, we screened for activation of DDR by immunoblotting of p-ATM, p-ATR, p-CHK1, p-CHK2 and p-TP53. Results: Compared to γH2AX foci levels in normal bone marrow samples (0.18 focus per CD34+ ± 0.6), increased levels of γH2AX foci were detected in 1/1 MDS cell line (6.4 foci per cell ± 0.0), 6/6 AML cell lines (12.0 foci per cell ± 0.6), 12/12 MDS bone marrow samples (2.8 foci per CD34+ ± 0.7) as well as 12/12 AML bone marrow samples (6.0 foci per blast ± 0.6). γH2AX and 53BP1 co-localized in all tested samples forming diffuse, clustered and marginal patterns. DDR proteins were expressed heterogeneously suggesting impairment of DDR. Conclusions: Our results provide evidence for a continuous increase of constitutive DSB across the spectrum from MDS to AML in the context of an impaired DDR. γH2AX and 53BP1 co-localize in unique patterns in nuclei of MDS and AML presumably owing to a non-random spatial organization of the genome and foci formation implies promotion of ineffective nonhomologous end-joining repair mechanisms at sites of constitutive DSB. Disclosures No relevant conflicts of interest to declare.

The pyrido[3,4-b]pyrazine systems investigated were found to form Reis- sert compounds in the presence of an acyl halide (or chloroformate) and trimethylsilyl cyanide. Only the mono Reissert compounds were isolated. In the case of 2,3-diphenylpyrido[3,4-b]pyrazine (7), its reaction with benzene sulfonyl chloride and trimethylsilyl cyanide gave 5-cyano-2,3- diphenylpyrido[3,4-b]pyrazine (16). Alkylation of the Reissert compound 11 and analog 12 either directly gave the alkylated hetercyclic base or the alkylated Reissert analog compound

This work details the benefits of ultra deep submicron (UDSM) SOI CMOS technology for high performance mixed signal circuits at mm-wave frequencies. In particular, a mm-wave Direct Digital Synthesizer (DDS) design in 45nm partially depleted (PD) SOI CMOS is presented with post extraction simulated performance of 32 Gsps sample rate that rivals state of the art III–V DDS performance. Technology benefits of the 45nm PD-SOI technology's billion transistor integration, sub-5pS digital gate delays, and measured ∼400GHz f t /∼200GHz f max cutoff frequency performance is highlighted. Increasing design challenges for UDSM CMOS mm-wave mixed signal circuits caused by increased gate leakage, limited transistor output impedance, inadequate foundry models, and required checking for design manufacturability are also addressed.

Genetic instability due to increased DNA damage and altered DNA repair is of central significance in the initiation and progression of inherited and sporadic human leukemias. Although very rare, some inherited DNA repair insufficiency syndromes (e.g., Fanconi anemia, Bloom's syndrome) have added substantially to our understanding of crucial mechanisms of leukemogenesis in recent years. Conversely, sporadic leukemias account for the main proportion of leukemias and here DNA damaging reactive oxygen species (ROS) play a central role. Although the exact mechanisms of increased ROS production remain largely unknown and no single pathway has been detected thus far, some oncogenic proteins (e.g., the activated tyrosine kinases BCR-ABL1 and FLT3-ITD) seem to play a key role in driving genetic instability by increased ROS generation which influences the disease course (e.g., blast crisis in chronic myeloid leukemia or relapse in FLT3-ITD positive acute myeloid leukemia). Of course other mechanisms, which promote genetic instability in leukemia also exist. A newly emerging mechanism is the genome-wide alteration of epigenetic marks (e.g., hypomethylation of histone H3K79), which promotes chromosomal instability. Taken together genetic instability plays a critical role both in inherited and sporadic leukemias and emerges as a common theme in both inherited and sporadic leukemias. Beyond its theoretical impact, the analysis of genetic instability may lead the way to the development of innovative therapy strategies.

The vertebrate skeletal muscle gives rise to a series of x-ray reflexions indexed as orders (n) of 77 nm, the even orders being meridional whereas the odd orders being near-meridional. The diffraction intensities associated with these reflexions originate from the axial period of 39 nm attributable to the repeat of troponin-tropomyosin on the thin filament. In the present study, the x-ray intensities of the furthest inner reflexions, A2 (n = 2) reflexion at an axial spacing of 1/39 nm-1 and A4 (n = 4) reflexion at 1/19 nm, of this series were measured with a time resolved manner. Upon activation of the frog striated muscle, the two reflexions underwent biphasic time courses of the intensity changes. With A2 reflexion, a rapid intensity increase by 16%, being completed by the time when tension rises to 5%, was followed by a slow intensity decrease down to 50%, which was associated with the tension rise. In both phases, lateral widths remained unchanged. A4 reflexion also behaves in the same way, although the first phase (the intensity increase) was not clear due to unsatisfactory statistics. We interpret phase one as being caused by conformational change of the troponin-tropomyosin complex upon binding of Ca2+ to troponin, whereas phase two being due to direct contribution of the mass of the myosin heads bound to the thin filament, although possible contribution of conformational changes of the regulatory proteins to phase two is not excluded. The results indicated that the calcium activation of the thin filament leads the onset of the actomyosin interaction.

Reissert compounds derived from α,β-unsaturated acid chlorides were prepared. The conjugate base obtained from these Reissert compounds exhibited the following carbanion reactions: 1) Alkylation, 2) Condensation with benzaldehyde, 3) Rearrangement to give dimeric compounds rather than simple rearranged compounds. In the case of alkylated isoquinoline Reissert compounds, the attempted rearrangement led to ring annellated amines.

In the presence of trimethylsilyl trifluoromethanesulfonate, a methylene chloride solution of isoquinoline and benzoyl chloride gave N-benzoylisoquinolinium triflate (3) and N-H-isoquinolinium triflate (4). Depending on the reaction conditions, reaction of 3 may occur on the isoquinoline ring yielding a Reissert compound and 1,2-dihydroisoquinoline species. Otherwise, reactions transpire on the carbonyl of 3 to give an amide, an ester, and an anhydride.

The FIP1L1-PDGFRA fusion gene has been described in patients with eosinophilia-associated myeloproliferative disorders (Eos-MPD). Here, we report on seven FIP1L1-PDGFRA-positive patients who presented with acute myeloid leukemia (AML, n=5) or lymphoblastic T-cell non-Hodgkin-lymphoma (n=2) in conjunction with AML or Eos-MPD. All patients were male, the median age was 58 years (range, 40-66). AML patients were negative for common mutations of FLT3, NRAS, NPM1, KIT, MLL and JAK2; one patient revealed a splice mutation of RUNX1 exon 7. Patients were treated with imatinib (100 mg, n=5; 400 mg, n=2) either as monotherapy (n=2), as maintenance treatment after intensive chemotherapy (n=3) or in overt relapse 43 and 72 months, respectively, after primary diagnosis and treatment of FIP1L1-PDGFRA-positive disease (n=2). All patients are alive, disease-free and in complete hematologic and complete molecular remission after a median time of 20 months (range, 9-36) on imatinib. The median time to achievement of complete molecular remission was 6 months (range, 1-14). We conclude that all eosinophilia-associated hematological malignancies should be screened for the presence of the FIP1L1-PDGFRA fusion gene as they are excellent candidates for treatment with tyrosine kinase inhibitors even if they present with an aggressive phenotype such as AML.

Background S100A8 and S100A9, also known as myeloid-related protein-8 (MRP8) and MRP14, are widely used as biomarkers of disease activity in juvenile idiopathic arthritis. On a molecular level these proteins function as damage-associated molecular pattern molecules (DAMPs), which amplify inflammation via Toll-like receptor-4 (TLR-4) activation. Released from activated phagocytes or necrotic cells the physiologically relevant S100A8/A9 heterodimers stimulate surrounding cells in order to promote pro-inflammatory processes in innate immunity. Adaptive immunity is believed to be enhanced by these molecules via activation of dendritic cells (DCs). Interestingly, recent studies show an additional regulatory effect of S100 proteins on DCs upon prolonged exposure (Petersen et al. EMBO-J 2013). Objectives The aim of this study is to decipher the S100 protein dependent molecular mechanisms in dendritic cells which enforce aggravation or attenuation of inflammation. Methods Human monocyte derived or murine bone marrow derived DCs are differentiated with or without exposure to S100A8 for six days prior to activation with LPS. After characterization and determination of the activation status using flow cytometry, the ability of these cells to induce T-cell proliferation is investigated in an autologous, antigen-specific or allogenic fashion. To identify the molecular mechanisms leading to the observed phenotype the mRNA expression of human DCs in various stages of differentiation was screened by genome-wide gene expression arrays. Results In a model of allergic contact dermatitis S100A9 deficient mice developed elevated inflammatory responses compared to wild type mice. We could demonstrate that prolonged exposure of myeloid progenitor cells to S100 proteins blocks DC differentiation and antigen presentation resulting in a reduced OT1 and OT2 T-cell response. Human S100A8/A9 shows a similar regulatory impact on monocyte-derived DCs (moDCs), since early DC differentiation and subsequent antigen-presentation is inhibited by S100A8 as well, leading to altered T-cell responses associated with decreased proliferation and enhanced IL-10 secretion. Gene expression analysis of developing DCs indicates vast changes in prominent immune modulatory pathways like JAK/STAT, NOD, RIG-I and TLR signalling. In addition, cell adhesion and antigen presentation seems to be affected by metabolic changes (e.g. PPAR-γ regulated fatty acid biosynthesis) as well as surface expression of antigen presenting molecules like HLA and CD1c. Conclusions Taken together, our results represent a novel regulatory mechanism of innate immunity to prevent overwhelming immune responses. Disclosure of Interest None declared

SRI International has prepared dynamically generated silica samples since 1980 for the National Institute of Occupational Safety and Health (NIOSH) and the American Industrial Hygiene Association (AIHA) Proficiency Analytical Testing (PAT) programs. Aerosol-generated samples were developed in 1980 to more closely approximate real world samples and to improve intrabatch precision. Liquid-generated samples may provide tighter control limits, and this method has been reexamined as the generation procedure of choice. Sample preparation procedures have also been investigated to minimize analytical uncertainty and, hence, obtain a true evaluation of the sampling error. Samples were analyzed by SRI, NIOSH, and the Wisconsin Occupational Health Laboratory, using X-ray diffraction or Fourier transform infrared spectrometry (FTIR). Results were plotted and statistically evaluated, then compared to the existing PAT interlaboratory database.