BTLA mediates inhibition of human tumor-specific CD8+ T cells that can be partially reversed by vaccination.

The function of antigen-specific CD8+ T cells, which may protect against both infectious and malignant diseases, can be impaired by ligation of their inhibitory receptors, which include CTL-associated protein 4 (CTLA-4) and programmed cell death 1 (PD-1). Recently, B and T lymphocyte attenuator (BTLA) was identified as a novel inhibitory receptor with structural and functional similarities to CTLA-4 and PD-1. BTLA triggering leads to decreased antimicrobial and autoimmune T cell responses in mice, but its functions in humans are largely unknown. Here we have demonstrated that as human viral antigen-specific CD8+ T cells differentiated from naive to effector cells, their surface expression of BTLA was gradually downregulated. In marked contrast, human melanoma tumor antigen-specific effector CD8+ T cells persistently expressed high levels of BTLA in vivo and remained susceptible to functional inhibition by its ligand herpes virus entry mediator (HVEM). Such persistence of BTLA expression was also found in tumor antigen-specific CD8+ T cells from melanoma patients with spontaneous antitumor immune responses and after conventional peptide vaccination. Remarkably, addition of CpG oligodeoxynucleotides to the vaccine formulation led to progressive downregulation of BTLA in vivo and consequent resistance to BTLA-HVEM-mediated inhibition. Thus, BTLA activation inhibits the function of human CD8+ cancer-specific T cells, and appropriate immunotherapy may partially overcome this inhibition.

Assessment of high hydrostatic pressure and starter culture on the quality properties of low-acid fermented sausages

The addition of starter culture and high pressure processing after ripening improved the microbial quality of low-acid fermented sausages (fuet and chorizo). The use of Lactobacillus sakei CTC6626 and Staphylococcus xylosus CTC6013 as starter culture significantly reduced Enterobacteriaceae and Enterococcus levels in the finished sausages. Moreover, the addition of starter culture produced sausages of similar quality to traditional low-acid fermented sausages. Slightly lower pH values and higher cohesiveness were obtained for both fuet and chorizo with starter culture. Sensory analysis showed no differences between lots of chorizo whereas starter fuet was more acid and gummy. High pressure induced an additional reduction of Enterobacteriaceae in non-starter sausages. An increase of textural properties was observed after pressurization. No other differences were observed between non-treated and pressurized sausages.

Combined effect of natural antimicrobials and high pressure processing to prevent Listeria monocytogenes growth after a cold chain break during storage of cooked ham

The effect of high pressure processing (400 MPa for 10 min) and natural antimicrobials 2 (enterocins and lactate-diacetate) on the behaviour of L. monocytogenes in sliced cooked ham 3 during refrigerated storage (1ºC and 6ºC) was assessed. The efficiency of the treatments after a 4 cold chain break was evaluated. Lactate-diacetate exerted a bacteriostatic effect against L. 5 monocytogenes during the whole storage period (3 months) at 1ºC and 6ºC, even after 6 temperature abuse. The combination of low storage temperature (1ºC), high pressure 7 processing (HPP) and addition of lactate-diacetate reduced the levels of L. monocytogenes 8 during storage by 2.7 log CFU/g. The most effective treatment was the combination of HPP, 9 enterocins and refrigeration at 1ºC, which reduced the population of the pathogen to final counts 10 of 4 MPN/g after 3 months of storage, even after the cold chain break.

Is chemotherapy necessary for premenopausal women with lower-risk node-positive, endocrine responsive breast cancer? 10-year update of International Breast Cancer Study Group Trial 11-93.

INTRODUCTION: International Breast Cancer Study Group (IBCSG) Trial 11-93 is the largest trial evaluating the role of the addition of chemotherapy to ovarian function suppression/ablation (OFS) and tamoxifen in premenopausal patients with endocrine-responsive early breast cancer. METHODS: IBCSG Trial 11-93 is a randomized trial comparing four cycles of adjuvant chemotherapy (AC: doxorubicin or epirubicin, plus cyclophosphamide) added to OFS and 5 years of tamoxifen versus OFS and tamoxifen without chemotherapy in premenopausal patients with node-positive, endocrine-responsive early breast cancer. There were 174 patients randomized from May 1993 to November 1998. The trial was closed before the target accrual was reached due to low accrual rate. RESULTS: Patients randomized tended to have lower risk node-positive disease and the median age was 45. After 10 years median follow up, there remains no difference between the two randomized treatment groups for disease-free (hazard ratio=1.02 (0.57-1.83); P=0.94) or overall survival (hazard ratio=0.97 (0.44-2.16); P=0.94). CONCLUSION: This trial, although small, offers no evidence that AC chemotherapy provides additional disease control for premenopausal patients with lower-risk node-positive endocrine-responsive breast cancer who receive adequate adjuvant endocrine therapy. A large trial is needed to determine whether chemotherapy adds benefit to endocrine therapy for this population.

Empirical use of anti-Gram-positive antibiotics in febrile neutropaenic cancer patients with acute leukaemia

Gram-positive infections including those due to methicillin-resistant staphylococci occur frequently in febrile neutropaenic patients. Although few data support the empirical addition of a glycopeptide antibiotic to the standard broad-spectrum antibiotic regimen, these agents are often used in many cancer centres. The emergence of infections due to vancomycin- resistant enterococci and glycopeptide-intermediate staphylococci has led to recommendations for a restricted use of glycopeptide antibiotics. The objective of the present work was to formulate evidence-based guidelines for the empirical use of anti- Gram-positive antibiotics in neutropaenic patients with acute leukaemia.

Marked increase of venlafaxine enantiomer concentrations as a consequence of metabolic interactions: a case report.

On three occasions, unusually high trough plasma concentrations of venlafaxine were measured in a patient phenotyped and genotyped as being an extensive CYP2D6 metabolizer and receiving 450 mg/day of venlafaxine and multiple comedications. Values of 1.54 and of 0.60 mg/l of venlafaxine and O-desmethylvenlafaxine, respectively, were determined in the first blood sample, giving an unusually high venlafaxine to O-desmethylvenlafaxine ratio. This suggests an impaired metabolism of venlafaxine to O-desmethylvenlafaxine, and is most likely due to metabolic interactions with mianserin (240 mg/day) and propranolol (40 mg/day). Concentration of (S)-venlafaxine measured in this blood sample was almost twice as high as (R)-venlafaxine ((S)/(R) ratio: 1.94). At the second blood sampling, after addition of thioridazine (260 mg/day), which is a strong CYP2D6 inhibitor, concentrations of venlafaxine were further increased (2.76 mg/l), and concentrations of O-desmethylvenlafaxine decreased (0.22 mg/l). A decrease of the (S)/(R)-venlafaxine ratio (-20%) suggests a possible stereoselectivity towards the (R)-enantiomer of the enzyme(s) involved in venlafaxine O-demethylation at these high venlafaxine concentrations. At the third blood sampling, after interruption of thioridazine, concentrations of venlafaxine and O-desmethylvenlafaxine were similar to those measured in the first blood sample. This case report shows the importance of performing studies on the effects of either genetically determined or acquired deficiency of metabolism on the kinetics of venlafaxine.

Performance of Holstein heifers fed sugarcane silages treated with urea, sodium benzoate or Lactobacillus buchneri

The objective of this work was to evaluate the performance of heifers fed sugarcane silages produced with and without additives. Thirty-two Holstein heifers were randomly assigned, in a block design, to evaluate rations (46% silage; 54% concentrate; 12% crude protein) containing silages treated with (fresh basis) urea (0.5%), sodium benzoate (0.1%) or Lactobacillus buchneri (3.64x10(5) cfu g-1 ). Inoculation with L. buchneri improved daily gain (1.24 vs. 0.94 kg day-1 ), and the addition of benzoate resulted in better feed conversion (7.6 vs. 9.4 kg of dry matter per kg of live weight), in relation to the untreated silage (control). Treatments did not affect dry matter intake (mean of 2.19% of live weight). Rations containing silages treated with benzoate or L. buchneri showed lower cost per kg of weight gain. Treatment with urea did not improve animal performance, but the cost per kg of weight gain was lower than that of the control ration.

Field Evaluation of Bonded Concrete Overlays, HR-288, 1986

A field program of strain and deflection measurements was conducted by the Construction Technology Laboratories (CTL) for the Iowa Department of Transportation. The objective of the field measurement program was to obtain information on bonded concrete resurfaced pavements that can be used as a data base for verifying bonded resurfacing thickness design procedures. Data gathered during the investigation included a visual condition survey, engineering properties of the original and resurfacing concrete, load related strain and deflection measurements, and temperature related curl (deflection) measurements. Resurfacing is basically the addition of a surface layer to extend the life of an existing pavement. Portland cement concrete has been used to resurface existing pavements since about 1913.

Evidence for a new post-translational modification in Staphylococcus aureus: Hydroxymethylation of asparagine and glutamine.

Staphylococcus aureus is an opportunistic pathogen whose infectious capacity depends on surface proteins, which enable bacteria to colonize and invade host tissues and cells. We analyzed "trypsin-shaved" surface proteins of S. aureus cultures by high resolution LC-MS/MS at different growth stages and culture conditions. Some modified peptides were identified, with a mass shift corresponding to the addition of a CH(2)O group (+30.0106u). We present evidence that this shift corresponds to a hyxdroxymethylation of asparagine and glutamine residues. This known but poorly documented post-translational modification was only found in a few proteins of S. aureus grown under specific conditions. This specificity seemed to exclude the hypothesis of an artifact due to sample preparation. Altogether hydroxymethylation was observed in 35 peptides from 15 proteins in our dataset, which corresponded to 41 modified sites, 35 of them being univocally localized. While no function can currently be assigned to this post-translational modification, we hypothesize that it could be linked to modulation of virulence factors, since it was mostly found on some surface proteins of S. aureus.

Strength Development of Concrete Based on Maturity and Pulse Velocity, MLR-95-03, 1995

The effect of curing temperature, in the range of 4.4 to 22.8 degrees C (40 to 73 degrees F), on strength development was studied based on the maturity and pulse velocity measurements in this report. The strength-maturity relationships for various mixes using a Type I cement and using a Type IP cement, respectively, were experimentally developed. The similar curves for early age strength development of both the patching concrete, using a Type I cement with the addition of calcium chloride, and the fast track concrete, using a Type III cement and fly ash, have also been proposed. For the temperature ranges studied, the strength development of concrete can be determined using a pulse velocity measurement, but only for early ages up to 24 hours. These obtained relationships can be used to determine when a pavement can be opened to traffic. The amount of fly ash substitution, up to 30%, did not have a significant influence on the strength-maturity relationship.

Physical performance of biodegradable films intended for antimicrobial food packaging

Antimicrobial films were prepared by including enterocins to alginate, polyvinyl alcohol (PVOH), and zein films. The physical performance of the films was assessed by measuring color, microstructure (SEM), water vapor permeability (WVP), and tensile properties. All studied biopolymers showed poor WVP and limited tensile properties. PVOH showed the best performance exhibiting the lowest WVP values, higher tensile properties, and flexibility among studied biopolymers. SEM of antimicrobial films showed increased presence of voids and pores as a consequence of enterocin addition. However, changes in microstructure did not disturb WVP of films. Moreover, enterocin-containing films showed slight improvement compared to control films. Addition of enterocins to PVOH films had a plasticizing effect, by reducing its tensile strength and increasing the strain at break. The presence of enterocins had an important effect on tensile properties of zein films by significantly reducing its brittleness. Addition of enterocins, thus, proved not to disturb the physical performance of studied biopolymers. Development of new antimicrobial biodegradable packaging materials may contribute to improving food safety while reducing environmental impact derived from packaging waste.

Functional expression of PHO1 to the Golgi and trans-Golgi network and its role in export of inorganic phosphate.

Arabidopsis thaliana PHO1 is primarily expressed in the root vascular cylinder and is involved in the transfer of inorganic phosphate (Pi) from roots to shoots. To analyze the role of PHO1 in transport of Pi, we have generated transgenic plants expressing PHO1 in ectopic A. thaliana tissues using an estradiol-inducible promoter. Leaves treated with estradiol showed strong PHO1 expression, leading to detectable accumulation of PHO1 protein. Estradiol-mediated induction of PHO1 in leaves from soil-grown plants, in leaves and roots of plants grown in liquid culture, or in leaf mesophyll protoplasts, was all accompanied by the specific release of Pi to the extracellular medium as early as 2-3 h after addition of estradiol. Net Pi export triggered by PHO1 induction was enhanced by high extracellular Pi and weakly inhibited by the proton-ionophore carbonyl cyanide m-chlorophenylhydrazone. Expression of a PHO1-GFP construct complementing the pho1 mutant revealed GFP expression in punctate structures in the pericycle cells but no fluorescence at the plasma membrane. When expressed in onion epidermal cells or in tobacco mesophyll cells, PHO1-GFP was associated with similar punctate structures that co-localized with the Golgi/trans-Golgi network and uncharacterized vesicles. However, PHO1-GFP could be partially relocated to the plasma membrane in leaves infiltrated with a high-phosphate solution. Together, these results show that PHO1 can trigger Pi export in ectopic plant cells, strongly indicating that PHO1 is itself a Pi exporter. Interestingly, PHO1-mediated Pi export was associated with its localization to the Golgi and trans-Golgi networks, revealing a role for these organelles in Pi transport.

The Utilization of Agriculturally Derived Lignin as an Antioxidant in Asphalt Binder, TR-557, 2008

Oxidation is the primary cause of long-term aging in asphalt pavements. As a pavement oxidizes, it stiffens and can eventually crack. The use of an antioxidant as a performance enhancer in an asphalt binder could delay aging, thus increasing the life of an asphalt pavement. Lignin is a highly available and well-studied antioxidant. A wet-mill ethanol plant produces several co-products, some of which contain lignin. The use of lignin from ethanol production could provide a benefit to asphalt pavements and also give more value to the co-products. The following research examined the effects of lignin on asphalt pavements. Three lignin-containing co-products were separately combined with four asphalt binders in varying amounts to determine the optimum amount of co-product that would provide the greatest benefit to the asphalt binders. The asphalt binder and co-product blends were evaluated according to Superpave specifications and performance graded on a continuous scale. The data indicated a stiffening effect on the binder caused by the addition of the co-products. The more a co-product was added, the more a binder stiffened. Binder stiffening benefited the high temperature properties and the low temperature binder properties were negatively affected. However, the low temperature stiffening effects were small and in many cases not significant. The co-products had an overall effect of widening the temperature range of the binders. This result suggests some antioxidant activity between the binder and the lignin. Testing with a fourth co-product with no lignin supported the idea that lignin acts as an antioxidant. The samples with no lignin aged significantly more than the samples with lignin. Infrared spectrometry also supported the idea that lignin acts as an antioxidant by observing decreases in some oxidative aging products.

Evaluation of Dowel Bar Retrofits for Local Road Pavements, TR-520, 2008

As truck traffic on Iowa secondary roads has increased, engineers have moved to concrete pavements of greater depths. Early designs included thickened edge pavements and depths of seven inches or greater. The designs typically did not have load transfer devices installed in the transverse joints and relied on aggregate interlock for this purpose. In some cases, aggregate interlock was not adequate to deal with the soils and traffic conditions and faulting of the joints has begun to appear. Engineers are now faced with the need to install or retrofit load transfer in the joints to preserve the pavements. Questions associated with this decision range from the type of dowel material to dowel diameter, spacing, number of bars, placement method, and construction techniques to be used to assure reduction or elimination of faulting. Buena Vista County constructed a dowel bar retrofit project on one mile of road. The plan called for addition of the dowels (2, 3, or 4) in the outer wheel path only and surface grinding in lieu of asphalt overlay. The project included the application of elliptical- and round-shaped dowels in a rehabilitation project. Dowel material types included conventional epoxy-coated steel and fiber-reinforced polymer (FRP). This work involved the determination of relative costs in materials to be used in this type of work and performance of FRP and elliptical-shaped steel dowels in the retrofit work. The results indicate good performance from each of the bar configurations and use the results of ride and deflection testing over the research period to project the benefits that can be gained from each configuration vs. the anticipated construction costs. The reader is cautioned that this project could not relate the number of dowels required to the level of anticipated truck traffic for other roads that might be considered.

Evaluation of High-Slump Concrete for Bridge Deck Overlays, TR-427, 2005

The Iowa Method for bridge deck overlays has been very successful in Iowa since its adoption in the 1970s. This method involves removal of deteriorated portions of a bridge deck followed by placement of a layer of dense (Type O) Portland Cement Concrete (PCC). The challenge encountered with this type of bridge deck overlay is that the PCC must be mixed on-site, brought to the placement area and placed with specialized equipment. This adds considerably to the cost and limits contractor selection, because not all contractors have the capability or equipment required. If it is possible for a ready-mix supplier to manufacture and deliver a dense PCC to the grade, then any competent bridge deck contractor would be able to complete the job. However, Type O concrete mixes are very stiff and generally cannot be transported and placed with ready-mix type trucks. This is where a “super-plasticizer” comes in to use. Addition of this admixture provides a substantial increase in the workability of the concrete – to the extent that it can be delivered to the site and placed on the deck directly out of a ready-mix truck. The objective of this research was to determine the feasibility of placing a deck overly of this type on county bridges within the limits of county budgets and workforce/contractor availability.