A recyclable assay to analyze the NH(2)-terminal trimming of antigenic peptide precursors.

The proteasome plays an essential role in the production of MHC class I-restricted antigenic peptides. Recent results have indicated that several peptidases, including tripeptidyl peptidase II and puromycin-sensitive aminopeptidase, could act downstream of the proteasome by trimming NH(2)-terminal extensions of antigenic peptide precursors liberated by the proteasome. In this study, we have developed a solid-phase peptidase assay that allowed us to efficiently purify and immobilize proteasome, tripeptidyl peptidase II, and puromycin-sensitive aminopeptidase. Whereas the first peptidase was active against small fluorogenic peptides, the latter two could also digest antigenic peptide precursors and could be used repeatedly with different precursors. Using three distinct antigenic peptide precursors, we found that tripeptidyl peptidase II never cleaved within the antigenic peptide sequence, suggesting that, aside from its proteolytic activities, it may also play a role in protecting antigenic peptides from complete hydrolysis in the cytosol. This method should be valuable for high throughput screenings of substrate specificity and potential inhibitors.

Adjuvant radioimmunotherapy trial with iodine-131-labeled anti-carcinoembryonic antigen monoclonal antibody F6 F(ab')2 after resection of liver metastases from colorectal cancer.

PURPOSE: To evaluate the feasibility of radioimmunotherapy (RIT) with radiolabeled anti-carcinoembryonic antigen antibodies after complete resection of liver metastases (LM) from colorectal cancer. Patients and Methods: Twenty-two patients planned for surgery of one to four LM received a preoperative diagnostic dose of a 131I-F(ab')2-labeled anti-carcinoembryonic antigen monoclonal antibody F6 (8-10 mCi/5 mg). 131I-F(ab')2 uptake was analyzed using direct radioactivity counting, and tumor-to-normal liver ratios were recorded. Ten patients with tumor-to-normal liver ratios of >5 and three others were treated with a therapeutic injection [180-200 mCi 131I/50 mg F(ab')2] 30 to 64 days after surgery. RESULTS: Median 131I-F(ab')2 immunoreactivity in patient serum remained at 91% of initial values for up to 96 hours after injection. The main and dose-limiting-toxicity was hematologic, with 92% and 85% grades 3 to 4 neutropenia and thrombocytopenia, respectively. Complete spontaneous recovery occurred in all patients. No human anti-mouse antibody response was observed after the diagnosis dose; however, 10 of the 13 treated patients developed human anti-mouse antibody approximately 3 months later. Two treated patients presented extrahepatic metastases at the time of RIT (one bone and one abdominal node) and two relapsed within 3 months of RIT (one in the lung and the other in the liver). Two patients are still alive, and one of these is disease-free at 93 months after resection. At a median follow-up of 127 months, the median disease-free survival is 12 months and the median overall survival is 50 months. CONCLUSION: RIT is feasible in an adjuvant setting after complete resection of LM from colorectal cancer and should be considered for future trials, possibly in combination with chemotherapy, because of the generally poor prognosis of these patients.

Assessment of a two-step nucleic acid amplification assay for detection of Neisseria meningitidis followed by capsular genogrouping

Immediate prevention of meningococcal disease relies in part on the prompt treatment with antibiotics of household and other close contacts of cases; however intervention with effective vaccination relies on identification of serogroup-causing strains. Parenteral antibiotic for patient with suspected meningococcal disease before hospital admission is currently recommended. Laboratory standard methods are hindered by failure to detect bacteria by this medical approach to improve patient prognosis. We assessed two polymerase chain reaction (PCR) assays to detect (crgA) and define the serogroups (siaD, orf-2, and ctrA) of Neisseria meningitidis in 120 cerebrospinal fluid (CSF) samples from positive cases (culture or antigen detection or direct smear). The PCR sensitivity for the identification of N. meningitidis was 100% (95% confidence interval, CI, 96-100%) compared to a sensitivity of 46% for culture (95% CI 37-55%), 61% for latex agglutination test (95% CI 52-70%), and 68% for Gram stain (95% CI 59-76%); PCR specificity was 97% (95% CI 82-100%). PCR correctly identified the serogroups A, B, C, W135, Y, and X in CSF samples with a sensitivity of 88% (95% CI 80-93%); the primer sets were 100% specific. The introduction of PCR-based assays shall increase laboratory confirmed cases, consequently enhancing surveillance of meningococcal disease.

Interactions between grain size and composition of sediments: two examples

Two contrasting case studies of sediment and detrital mineral composition are investigated in order to outline interactions between chemical composition and grain size. Modern glacial sediments exhibit a strong dependence of the two parameters due to the preferential enrichment of mafic minerals, especially biotite, in the fine-grained fractions. On the other hand, the composition of detrital heavy minerals (here: rutile) appears to be not systematically related to grain-size, but is strongly controlled by location, i.e. the petrology of the source rocks of detrital grains. This supports the use of rutile as a well-suited tracer mineral for provenance studies. The results further suggest that (i) interpretations derived from whole-rock sediment geochemistry should be flanked by grain-size observations, and (ii) a more sound statistical evaluation of these interactions require the development of new tailor-made statistical tools to deal with such so-called two-way compositions

Immunostimulatory property of a synthetic peptide belonging to the soluble ATP diphosphohydro-lase isoform (SmATPDase 2) and immunolocalisation of this protein in the Schistosoma mansoni egg

A peptide (SmB2LJ; r175-194) that belongs to a conserved domain from Schistosoma mansoni SmATPDase 2 and is shared with potato apyrase, as predicted by in silico analysis as antigenic, was synthesised and its immunostimulatory property was analysed. When inoculated in BALB/c mice, this peptide induced high levels of SmB2LJ-specific IgG1 and IgG2a subtypes, as detected by enzyme linked immunosorbent assay. In addition, dot blots were found to be positive for immune sera against potato apyrase and SmB2LJ. These results suggest that the conserved domain r175-194 from the S. mansoni SmATPDase 2 is antigenic. Western blots were performed and the anti-SmB2LJ antibody recognised in adult worm (soluble worm antigen preparation) or soluble egg antigen antigenic preparations two bands of approximately 63 and 55 kDa, molecular masses similar to those predicted for adult worm SmATPDase 2. This finding strongly suggests the expression of this same isoform in S. mansoni eggs. To assess localisation of SmATPDase 2, confocal fluorescence microscopy was performed using cryostat sections of infected mouse liver and polyclonal antiserum against SmB2LJ. Positive reactions were identified on the external surface from the miracidium in von Lichtenberg's envelope and, in the outer side of the egg-shell, showing that this soluble isoform is secreted from the S. mansoni eggs.

Historia morbi atrocis--deux nouveaux cas de ruptures spontanées de l'oesophage (syndrome de Boerhaave) [Historia morbi atrocis--2 new cases of spontaneous rupture of the esophagus (Boerhaave syndrome)].]

We report the case of two patients hospitalized within a few weeks of each other and both presenting with spontaneous rupture of the esophagus whose evolution proved fatal. We take the opportunity of drawing attention to this rare and challenging disease, which is often diagnosed too late.

Estimation of electronic coupling in π -stacked donor-bridge-acceptor systems: correction of the two-state model

Comparison of donor-acceptor electronic couplings calculated within two-state and three-state models suggests that the two-state treatment can provide unreliable estimates of Vda because of neglecting the multistate effects. We show that in most cases accurate values of the electronic coupling in a π stack, where donor and acceptor are separated by a bridging unit, can be obtained as Ṽ da = (E2 - E1) μ12 Rda + (2 E3 - E1 - E2) 2 μ13 μ23 Rda2, where E1, E2, and E3 are adiabatic energies of the ground, charge-transfer, and bridge states, respectively, μij is the transition dipole moments between the states i and j, and Rda is the distance between the planes of donor and acceptor. In this expression based on the generalized Mulliken-Hush approach, the first term corresponds to the coupling derived within a two-state model, whereas the second term is the superexchange correction accounting for the bridge effect. The formula is extended to bridges consisting of several subunits. The influence of the donor-acceptor energy mismatch on the excess charge distribution, adiabatic dipole and transition moments, and electronic couplings is examined. A diagnostic is developed to determine whether the two-state approach can be applied. Based on numerical results, we showed that the superexchange correction considerably improves estimates of the donor-acceptor coupling derived within a two-state approach. In most cases when the two-state scheme fails, the formula gives reliable results which are in good agreement (within 5%) with the data of the three-state generalized Mulliken-Hush model

Dynamics of the two-dimensional gonihedric spin model

In this paper, we study dynamical aspects of the two-dimensional (2D) gonihedric spin model using both numerical and analytical methods. This spin model has vanishing microscopic surface tension and it actually describes an ensemble of loops living on a 2D surface. The self-avoidance of loops is parametrized by a parameter ¿. The ¿=0 model can be mapped to one of the six-vertex models discussed by Baxter, and it does not have critical behavior. We have found that allowing for ¿¿0 does not lead to critical behavior either. Finite-size effects are rather severe, and in order to understand these effects, a finite-volume calculation for non-self-avoiding loops is presented. This model, like his 3D counterpart, exhibits very slow dynamics, but a careful analysis of dynamical observables reveals nonglassy evolution (unlike its 3D counterpart). We find, also in this ¿=0 case, the law that governs the long-time, low-temperature evolution of the system, through a dual description in terms of defects. A power, rather than logarithmic, law for the approach to equilibrium has been found.

A Continuous Span Aluminum Girder Concrete Deck Bridge - Part 2 of 2: Fatigue Tests of Aluminum Girders, Final Report, 1997

In 1957, the Iowa State Highway Commission, with financial assistance from the aluminum industry, constructed a 220-ft (67-m) long, four-span continuous, aluminum girder bridge to carry traffic on Clive Road (86th Street) over Interstate 80 near Des Moines, Iowa. The bridge had four, welded I-shape girders that were fabricated in pairs with welded diaphragms between an exterior and an interior girder. The interior diaphragms between the girder pairs were bolted to girder brackets. A composite, reinforced concrete deck served as the roadway surface. The bridge, which had performed successfully for about 35 years of service, was removed in the fall of 1993 to make way for an interchange at the same location. Prior to the bridge demolition, load tests were conducted to monitor girder and diaphragm bending strains and deflections in the northern end span. Fatigue testing of the aluminum girders that were removed from the end spans were conducted by applying constant-amplitude, cyclic loads. These tests established the fatigue strength of an existing, welded, flange-splice detail and added, welded, flange-cover plates and horizontal web plate attachment details. This part, Part 2, of the final report focuses on the fatigue tests of the aluminum girder sections that were removed from the bridge and on the analysis of the experimental data to establish the fatigue strength of full-size specimens. Seventeen fatigue fractures that were classified as Category E weld details developed in the seven girder test specimens. Linear regression analyses of the fatigue test results established both nominal and experimental stress-range versus load cycle relationships (SN curves) for the fatigue strength of fillet-welded connections. The nominal strength SN curve obtained by this research essentially matched the SN curve for Category E aluminum weldments given in the AASHTO LRFD specifications. All of the Category E fatigue fractures that developed in the girder test specimens satisfied the allowable SN relationship specified by the fatigue provisions of the Aluminum Association. The lower-bound strength line that was set at two standard deviations below the least squares regression line through the fatigue fracture data points related well with the Aluminum Association SN curve. The results from the experimental tests of this research have provided additional information regarding behavioral characteristics of full-size, aluminum members and have confirmed that aluminum has the strength properties needed for highway bridge girders.

Influence of misoprostol or prostaglandin E(2) for induction of labor on the incidence of pathological CTG tracing: a randomized trial.

OBJECTIVE: To compare the efficacy and safety of misoprostol (prostaglandin E(1) (PGE(1))) with dinoprostone (prostaglandin E(2) (PGE(2))) for third trimester cervical ripening and labor induction. STUDY DESIGN: Patients requiring induction of labor were randomly assigned to receive either 50 microg of intravaginal misoprostol every 4 h or 0.5 mg of intracervical dinoprostone gel every 6 h. Eligibility criteria included gestation = 36 weeks. Primary outcome was the time interval from induction to delivery; secondary outcomes were mode of delivery, perinatal outcome, and interpretation of cardiotocogram (CTG) records. RESULTS: Two hundred women were randomly enrolled to receive either misoprostol (n = 100) or dinoprostone (n = 100). Time induction-to-delivery at 12, 24 and 48 h and the need for oxytocin were reduced with misoprostol (P < 0.05). Pathological CTG tracing according to FIGO and Melchior scores were more frequent in the misoprostol-treated group (P < 0.001). CONCLUSION: Misoprostol shortened the induction-to-delivery interval, but is associated with a higher incidence of abnormal CTG than prostaglandin E(2).

Resource-based approach to new product development at the intersection of industries. Two case studies in telecommunications

Työn tavoitteena on selvittää mitkä ovat tärkeimmät aineettomat resurssit, joita tarvitaan teollisuuksien risteyskohdassa tapahtuvassa tuotekehityksessä. Teollisuuksien risteyskohdissa syntyvät tuotteet ovat usein radikaaleja, mikä tekee tuotteista mielenkiintoisia, paljon liiketoimintapotentiaalia tarjoavia. Tämä tutkimus lähestyy tuotekehitystä resurssipohjaisesta näkökulmasta. Myös tietämyspohjaista ja suhdepohjaista näkemystä hyödynnetään korostamaan keskittymistä aineettomiin resursseihin. Tutkimuksessa rakennetaan viitekehys, jossa tutkitaan eri resurssikategorioita. Valitut kategoriat ovat teknologiset, markkinointi-, johtamiseen ja hallinnointiin liittyvät ja suhdepohjaiset resurssit. Empiirisessä osassa tutkitaan kahta uutta tuotekonseptia, jotka ovat syntyneet teollisuuksien risteyskohdissa. Empiirisen osan tavoitteena on määritellä tutkimuksen kohteena olevia alustavia tuotekonsepteja tarkemmin ja selvittää millaisia resursseja näiden toteuttamiseen tarvitaan. Myös tarvittavien resurssien nykytila selvitetään ja pohditaan tulisiko puuttuvia resursseja kehittää yrityksen sisällä vai hankkia ne ulkopuolelta. Tutkimus toteutettiin asiantuntijahaastatteluin. Kahden tapaustutkimuksen perusteella näyttäisi siltä, että suhdepohjaiset resurssit ovat erittäin tärkeitä teollisuuksien risteyskohdissa tapahtuvassa tuotekehityksessä. Myös teknologiset resurssit ovat tärkeitä. Markkinointiresurssien tärkeys riippuu lopullisesta tuotekonseptista, kun taas johtamiseen ja kehittämiseen liittyvät resurssit ovat tärkeitänäiden konseptien luomisessa.

Synthesis of the two isomers of the potential sex pheromone of Thaumetopoea pityocampa (Lepidoptera, Notodontidae) and related model compounds

The synthesis of the major component of the sex pheromone secretion of the processionary moth, Tkawnztopoeja pltyocampa (Denis and Schiff.) (Lepidoptera, Notodontidae), (Z)-13-hexadecen-ll-ynyl acetate (1), the corresponding (E)-isomer (2) and the four structurally related model compounds (Z⁄E,Z,Z)-5,9,13-hexadecatrienyl acetate (3), (Z⁄E,Z,Z)-3,7,ll-hexadecatrienyl acetate (4), (Z⁄E,E,Z)-7,9,13-hexadecatrienyl acetate (5) and (Z)-7-hexadecen-5-ynyl acetate (6) is described.

Elaborated Literary Violence: Genre and Ideology of the Two Stories I Sam 22,6-23 and II Sam 21,1-14

The present article focuses on two stories dealing with acts of big blood shed. I Sam 22,6-23 relates the massacre of the priests of Nob; II Sam 21,1-14 is about the execution of seven descendents of Saul, as atonement for Saul's attempt to exterminate the Gibeonites. Most researchers consider both stories or at least certain parts of them old.1 For both stories few verses are regarded as secondary. In this paper I try to reassess the analysis of these stories and will point to indices favoring a late date of origin for both of them. They concern the language in use, intertextual connections and particular motifs. A further indication consists in the fact that the reported events of the stories lack significant resonance in the corpus of the Hebrew Bible. There are only two texts, I Sam 2,33 and Ps 52,2, which allude or refer to I Sam 22,6-23.With regard to the importance of the related events and acts this silence in the Biblical context is astonishing. Interestingly, also in the Book of Chronicles one does not find any allusions to these stories. This raises the question whether the latter were composed after the formation of the book of Chronicles.

Nanoantenna enhanced emission of lightharvesting complex 2: the role of resonance, polarization, and radiative and non-radiative rates

Nanoantennae show potential for photosynthesis research for two reasons; first by spatially confining light for experiments which require high spatial resolution, and second by enhancing the photon emission of single light-harvesting complexes. For effective use of nanoantennae a detailed understanding of the interaction between the nanoantenna and the light-harvesting complex is required. Here we report how the excitation and emission of multiple purple bacterial LH2s (light-harvesting complex 2) are controlled by single gold nanorod antennae. LH2 complexes were chemically attached to such antennae, and the antenna length was systematically varied to tune the resonance with respect to the LH2 absorption and emission. There are three main findings. (i) The polarization of the LH2 emission is fully controlled by the resonant nanoantenna. (ii) The largest fluorescence enhancement, of 23 times, is reached for excitation with light at λ = 850 nm, polarized along the long antenna-axis of the resonant antenna. The excitation enhancement is found to be 6 times, while the emission efficiency is increased 3.6 times. (iii) The fluorescence lifetime of LH2 depends strongly on the antenna length, with shortest lifetimes of [similar]40 ps for the resonant antenna. The lifetime shortening arises from an 11 times resonant enhancement of the radiative rate, together with a 2–3 times increase of the non-radiative rate, compared to the off-resonant antenna. The observed length dependence of radiative and non-radiative rate enhancement is in good agreement with simulations. Overall this work gives a complete picture of how the excitation and emission of multi-pigment light-harvesting complexes are influenced by a dipole nanoantenna.