Why air quality in the Alps remains a matter of concern. The impact of organic pollutants in the alpine area.

In the middle of Europe, the Alps form a geographical and meteorological trap for atmospheric pollutants including volatile and semi-volatile organic compounds emitted in the surrounding lowlands. This is due to their barrier effects, high precipitation rates, and low ambient temperatures. Also the pollutants emitted in the cities inside the Alps spread in the region depending on orographic and meteorological conditions. Although a number of studies on the distribution and effect of pollutants in the Alps has been published, comprehensive information on potential hazards, and ways to improve this sensible environment are lacking. This opinion paper is the result of a discussion during the Winterseminar of the AlpsBioCluster project in Munich. It summarizes the current literature and presents some case studies on local pollution sources in the Alps, and the possibility of using biomonitoring techniques to assess critical pollution loads and distributions.

Ruminally undegradable protein content and digestibility for forages using the mobile bag in situ technique.

Four experiments were conducted to evaluate RUP content and digestibility for smooth bromegrass, subirrigated meadow, upland native range, and warm-season grasses. Samples were collected from esophageally cannulated cows or ruminally cannulated steers. Forages were ruminally incubated in in situ bags for durations of time based on 75% of total mean retention time, which was based on IVDMD and rate of passage calculations. One-half of the bags were duodenally incubated and excreted in the feces, and NDIN was analyzed on all bags for RUP calculations. Crude protein was numerically greater early in the growing cycle for grasses compared with later as grasses matured (P ≤ 0.32). The RUP was 13.3%, 13.3%, and 19.7% of CP for smooth bromegrass, subirrigated meadow, and upland native range, respectively. These values tended to be lower early in the growth cycle and increased (linear P ≤ 0.13) as forages matured for warm-season grasses and subirrigated meadows. Because both CP and RUP content change throughout the growing season, expressing RUP as a percentage of DM gives more consistent averages compared with RUP as a percentage of CP. Coefficient of variation values for RUP as a percentage of DM averaged 0.21 over all 4 experiments compared with 0.26 for RUP as a percentage of CP. Average RUP as a percentage of DM was 2.03%, 1.53%, and 1.94% for smooth bromegrass, subirrigated meadow, and upland native range, respectively. Total tract indigestible protein (TTIDP) linearly increased with maturity for subirrigated meadow samples (P < 0.01). A quadratic response (P ≤ 0.06) for TTIDP was observed in smooth bromegrass and warm-season grass samples. Digestibility of RUP varied considerably, ranging from 25% to 60%. Subirrigated meadow, native range, and smooth bromegrass samples tended to have linear decreases (P ≤ 0.11) in RUP digestibility throughout the growing season. The amount of digested RUP was fairly consistent across experiments and averages for smooth bromegrass, subirrigated meadow, and upland native range were 0.92%, 0.64%, and 0.49% of DM, respectively. Warm-season grasses in Exp. 2 had greater RUP (4.31% of DM) and amount of RUP digested (2.26% of DM), possibly because of cattle selecting for leadplant that contains more CP than the grasses. Forages can vary in CP, RUP, TTIDP, and RUP digestibility depending on the forage type, year, and time within year, but RUP digestibility is likely less than what previous sources have reported.

Contemporaneous cell spreading and phagocytosis: magneto-resistive real-time monitoring of membrane competing processes.

Adhesion and spreading of cells strongly depend on the properties of the underlying surface, which has significant consequences in long-term cell behavior adaption. This relationship is important for the understanding of both biological functions and their bioactivity in disease-related applications. Employing our magnetic lab-on-a-chip system, we present magnetoresistive-based real-time and label-free detection of cellular phagocytosis behavior during their spreading process on particle-immobilized sensor surfaces. Cell spreading experiments carried out on particle-free and particle-modified surfaces reveal a delay in spreading rate after an elapsed time of about 2.2h for particle-modified surfaces due to contemporaneous cell membrane loss by particle phagocytosis. Our associated magnetoresistive measurements show a high uptake rate at early stages of cell spreading, which decreases steadily until it reaches saturation after an average elapsed time of about 100 min. The corresponding cellular average uptake rate during the entire cell spreading process accounts for three particles per minute. This result represents a four times higher phagocytosis efficiency compared to uptake experiments carried out for confluently grown cells, in which case cell spreading is already finished and, thus, excluded. Furthermore, other dynamic cell-surface interactions at nano-scale level such as cell migration or the dynamics of cell attachment and detachment are also addressable by our magnetic lab-on-a-chip approach.

Magnetoresistive-based real-time cell phagocytosis monitoring.

The uptake of large particles by cells (phagocytosis) is an important factor in cell biology and also plays a major role in biomedical applications. So far, most methods for determining the phagocytic properties rely on cell-culture incubation and end-point detection schemes. Here, we present a lab-on-a-chip system for real-time monitoring of magnetic particle uptake by human fibroblast (NHDF) cells. It is based on recording the time evolution of the average position and distribution of magnetic particles during phagocytosis by giant-magnetoresistive (GMR) type sensors. We employ particles with a mean diameter of 1.2 µm and characterize their phagocytosis-relevant properties. Our experiments at physiological conditions reveal a cellular uptake rate of 45 particles per hour and show that phagocytosis reaches saturation after an average uptake time of 27.7h. Moreover, reference phagocytosis experiments at 4°C are carried out to mimic environmental or disease related inhibition of the phagocytic behavior, and our measurements clearly show that we are able to distinguish between cell-membrane adherent and phagocytosed magnetic particles. Besides the demonstrated real-time monitoring of phagocytosis mechanisms, additional nano-biointerface studies can be realized, including on-chip cell adhesion/spreading as well as cell migration, attachment and detachment dynamics. This versatility shows the potential of our approach for providing a multifunctional platform for on-chip cell analysis.

Transient lymphocyte decrease due to adhesion and migration following catumaxomab (anti-EpCAM x anti-CD3) treatment in vivo

INTRODUCTION: In patients, a transient decrease in peripheral blood lymphocyte counts was observed following intraperitoneal administration of the trifunctional monoclonal antibody catumaxomab (anti-human EpCAM x anti-human CD3). The aim of this study was to clarify the observed effect in a preclinical mouse model and to analyse the related mechanism of action in vitro. MATERIALS AND METHODS: A related antibody, BiLu (antihuman EpCAM x anti-mouse CD3), was administered to mice and blood leukocytes were analysed. In vitro studies measured activation and cytokine secretion from human peripheral blood mononuclear cells (PBMC). For the analysis of T cell adhesion, PBMC were preincubated with catumaxomab and then co-cultured with human endothelial cells (HUVEC); T cell adhesion was assessed in the presence or absence of endothelial cell preactivation by TNFα. Adherent T cells were determined by flow cytometry. RESULTS: Treatment of mice with BiLu resulted in a dosedependent transient decrease in CD3+ T cells (both CD4+ and CD8+) that returned to the normal range within 48 h. Catumaxomab physiologically activated T cells in vitro (increased CD69 expression) and induced cytokine release (TNFα, IFNγ). TNFα increased expression of adhesion molecules CD54 and CD62E on endothelial cells. Furthermore, catumaxomab dose-dependently enhanced adhesion of T cells to endothelial cells. Adhesion was further increased when endothelial cells were preactivated with TNFα. CONCLUSIONS: Catumaxomab increases adhesion of T cells to endothelial cells due to antibody-mediated activation of T cells and production of T cell cytokines that up-regulate endothelial cell adhesion molecules. These results provide a mechanistic rationale for the transient, reversible decrease in lymphocyte counts observed following catumaxomab administration in patients, which is likely to be due to redistribution of lymphocytes (AU)

Trifunctional antibodies induce efficient antitumour activity with immune cells from head and neck squamous cell carcinoma patients after radio-chemotherapy treatment.

BACKGROUND Trifunctional antibodies, such as catumaxomab (anti-EpCAM×anti-CD3) and ertumaxomab (anti- HER-2/neu×anti-CD3), transiently link immune effector cells to tumour cells, which results in cellular cytotoxicity towards the tumour cells. A functional immune system is therefore essential for effective anti-tumour activity. However, the commonly observed haematotoxicity of chemotherapeutics and radiation therapy may be associated with some degree of immunosuppression. Combining chemotherapy and trifunctional antibodies in cancer treatment requires understanding of the impact of chemotherapeutics on immune cell function and, thus, on the activity of trifunctional antibodies. METHODS The effect of chemotherapeutic treatment on trifunctional antibody-mediated anti-tumour activity was assessed in vitro. Blood samples were collected from 12 head and neck squamous cell carcinoma patients after chemotherapy (5-fluorouracil, cisplatin) and radiotherapy, and from one healthy control donor. The immune cell status was analysed and mononuclear cells (MNC) were isolated. The potency of catumaxomab and ertumaxomab was assessed in a cytotoxicity assay using MNC isolated from each patient sample in co-culture with a tumour target cell line. The release of infl ammatory cytokines was also monitored in the cell culture supernatant. RESULTS Most patients included in this study had decreased immune cell counts during the course of chemotherapy. Nonetheless, an effective and concentration-dependent anti- tumour activity mediated by trifunctional antibodies was demonstrated using these patient immune effector cells. The immune response activity of the patient immune cells was not impaired one week after cisplatin administration or even three days after the last 5-fluorouracil treatment. CONCLUSION This study shows for the first time that immune effector cells from cancer patients undergoing standard chemotherapy and radiotherapy can be activated by trifunctional antibodies for efficient killing of tumour cells.

Trifunctional antibodies induce efficient antitumour activity with immune cells from head and neck squamous cell carcinoma patients after radio-chemotherapy treatment

BACKGROUND Trifunctional antibodies, such as catumaxomab (anti-EpCAM×anti-CD3) and ertumaxomab (anti- HER-2/neu×anti-CD3), transiently link immune effector cells to tumour cells, which results in cellular cytotoxicity towards the tumour cells. A functional immune system is therefore essential for effective anti-tumour activity. However, the commonly observed haematotoxicity of chemotherapeutics and radiation therapy may be associated with some degree of immunosuppression. Combining chemotherapy and trifunctional antibodies in cancer treatment requires understanding of the impact of chemotherapeutics on immune cell function and, thus, on the activity of trifunctional antibodies. METHODS The effect of chemotherapeutic treatment on trifunctional antibody-mediated anti-tumour activity was assessed in vitro. Blood samples were collected from 12 head and neck squamous cell carcinoma patients after chemotherapy (5-fluorouracil, cisplatin) and radiotherapy, and from one healthy control donor. The immune cell status was analysed and mononuclear cells (MNC) were isolated. The potency of catumaxomab and ertumaxomab was assessed in a cytotoxicity assay using MNC isolated from each patient sample in co-culture with a tumour target cell line. The release of infl ammatory cytokines was also monitored in the cell culture supernatant. RESULTS Most patients included in this study had decreased immune cell counts during the course of chemotherapy. Nonetheless, an effective and concentration-dependent anti- tumour activity mediated by trifunctional antibodies was demonstrated using these patient immune effector cells. The immune response activity of the patient immune cells was not impaired one week after cisplatin administration or even three days after the last 5-fluorouracil treatment. CONCLUSION This study shows for the first time that immune effector cells from cancer patients undergoing standard chemotherapy and radiotherapy can be activated by trifunctional antibodies for efficient killing of tumour cells (AU)

Artificial cilia of magnetically tagged polymer nanowires for biomimetic mechanosensing.

Polymeric nanowires of polypyrrole have been implemented as artificial cilia on giant-magneto-resistive multilayer sensors for a biomimetic sensing approach. The arrays were tagged with a magnetic material, the stray field of which changes relative to the underlying sensor as a consequence of mechanical stimuli which are delivered by a piezoactuator. The principle resembles balance sensing in mammals. Measurements of the sensor output voltage suggest a proof of concept at frequencies of around 190 kHz and a tag thickness of ∼300 nm. Characterization was performed by scanning electron microscopy and magnetic force microscopy. Micromagnetic and finite-element simulations were conducted to assess basic sensing aspects.

What is needed for a sunscreen to provide complete protection.

Human skin is increasingly exposed to sunlight. In order to achieve complete protection against the cumulative detrimental effects from sun exposure, topical strategies must shield against the range of solar wavelengths that can damage the skin. Importantly, the harm sustained by the skin is not limited to that caused by the ultraviolet (UV) portion of the light spectrum, but also includes the adverse effects inflicted by near infrared energy. Consequently, in an attempt to provide the necessary broad spectrum coverage, innovative research continues through the exploration of new compounds and novel combinations of chemical and physical UV filters with molecules that are capable of interfering with and/or preventing the deleterious effects of infrared A (IRA) radiation. Existing examples of infrared-protective active agents include mitochondrially targeted antioxidants of synthetic or natural origin.

Photoprotection beyond ultraviolet radiation--effective sun protection has to include protection against infrared A radiation-induced skin damage.

Solar radiation is well known to damage human skin, for example by causing premature skin ageing (i.e. photoageing). We have recently learned that this damage does not result from ultraviolet (UV) radiation alone, but also from longer wavelengths, in particular near-infrared radiation (IRA radiation, 760-1,440 nm). IRA radiation accounts for more than one third of the solar energy that reaches human skin. While infrared radiation of longer wavelengths (IRB and IRC) does not penetrate deeply into the skin, more than 65% of the shorter wavelength (IRA) reaches the dermis. IRA radiation has been demonstrated to alter the collagen equilibrium of the dermal extracellular matrix in at least two ways: (a) by leading to an increased expression of the collagen-degrading enzyme matrix metalloproteinase 1, and (b) by decreasing the de novo synthesis of the collagen itself. IRA radiation exposure therefore induces similar biological effects to UV radiation, but the underlying mechanisms are substantially different, specifically, the cellular response to IRA irradiation involves the mitochondrial electron transport chain. Effective sun protection requires specific strategies to prevent IRA radiation-induced skin damage.

Prevention of infrared-A radiation mediated detrimental effects in human skin.

Photoaging and skin damage that is caused by solar radiation is well known. We have recently learned that within the solar spectrum this damage not only results from ultraviolet (UV) radiation, but also from longer wavelengths, in particular near infrared radiation. Accordingly, infrared radiation (IR) has been shown to alter the collagen equilibrium of the dermal extracellular matrix in at least 2 ways: (1) by leading to an increased expression of the collagen degrading enzyme matrixmetalloproteinase-1 while (2) decreasing the de novo synthesis of the collagen itself. Infrared-A (IRA) radiation exposure, therefore, induces similar biological effects to UV, but the underlying mechanisms are substantially different. IRA acts via the mitochondria and therefore protection from IR requires alternative strategies.

[IRA protection. Needs and possibilities]. / IRA-Schutz. Notwendigkeit und Möglichkeiten.

When it comes to skin damage, solar radiation is often regarded to be identical to ultraviolet (UV) but it includes much more. Over 90% of solar radiation is in the non-UV-range. Infrared A radiation (IRA, 760-1440 nm) accounts for around 30% of the solar energy reaching the earth's surface and exert detrimental effects on the skin. IRA alters the collagen equilibrium of the dermal extracellular matrix by leading to an increased expression of the collagen degrading enzyme matrixmetalloproteinase-1 while decreasing the expression of the dominant collagen gene Col1alpha1. IRA therefore leads to endpoints similar to UV, but the underlying biological mechanisms are substantially different. IRA acts via the mitochondria. IRA-specific protective approaches should be added to conventional sun protections strategies.

Assessment of the incremental value of recombinant thyrotropin stimulation before 2-[18F]-Fluoro-2-deoxy-D-glucose positron emission tomography/computed tomography imaging to localize residual differentiated thyroid cancer.

PURPOSE: The purpose of the study was to assess prospectively the impact of recombinant human TSH (rhTSH) administration on positron emission tomography (PET)/computed tomography (CT) imaging in differentiated thyroid cancer patients who, after primary treatment, had a suppressed or stimulated serum thyroglobulin greater than 10 ng/ml and no radioactive iodine uptake consistent with thyroid cancer on a whole body scan. PATIENTS AND METHODS: PET/CT was performed before (basal PET) and 24-48 h after rhTSH administration (rhTSH-PET) in 63 patients (52 papillary and 11 follicular thyroid cancers). Images were blindly analyzed by two readers. The proposed treatment plan was prospectively assessed before basal PET, after basal PET, and again after rhTSH-PET. RESULTS: A total of 108 lesions were detected in 48 organs in 30 patients. rhTSH-PET was significantly more sensitive than basal PET for the detection of lesions (95 vs. 81%; P = 0.001) and tended to be more sensitive for the detection of involved organs (94 vs. 79%; P = 0.054). However, basal PET and rhTSH-PET did not have significantly different sensitivity for detecting patients with any lesions (49 vs. 54%; P = 0.42). Changes in treatment management plan occurred in 19% of the patients after basal PET. Lesions found only by rhTSH-PET contributed to an altered therapeutic plan in eight patients, among whom only four were true-positive on pathology (6%). CONCLUSION: The use of rhTSH for 2-[18F]-fluoro-2-deoxy-D-glucose-PET/CT significantly increased the number of lesions detected, but the numbers of patients in whom any lesion was detected were no different between basal and rhTSH-stimulated PET/CT scans. Treatment changes due to true positive lesions occurred in 6% of cases.

The effect of processed water on constitutive and ultraviolet-A-radiation-induced level of mitochondrial DNA mutations in human dermal fibroblasts.

The integrity of the mitochondrial genome is of crucial importance for the cellular energy metabolism, and increased mutation rates are causally related to premature ageing. We demonstrate that replacement of normal deionized water with processed water in cell culture medium decreases the constitutive levels of the most frequent large-scale deletion of the mitochondrial genome in human dermal fibroblasts. In addition the presence of processed water also prevented the generation of the common deletion which was induced in these cells by repetitive UVA irradiation (3 x 8 J/cm(2) daily). Thus, processed water appears to protect the mitochondrial genome and may thus exert anti-oxidative and anti-ageing effects.

Education in end-of-life care: bridging disciplinary and institutional boundaries.

This paper describes the development of an interdisciplinary, interinstitutional seminar in palliative care for South Dakota students in medicine, nursing, pharmacy, chaplaincy, and social work. Student outcomes from six seminars conducted during 2001-2004 are reported, and recommendations for future educational efforts are outlined.

Rainfall timing and poultry litter application rate effects on phosphorus loss in surface runoff.

Phosphorus (P) in runoff from pastures amended with poultry litter may be a significant contributor to eutrophication of lakes and streams in Georgia and other areas in the southeastern United States. The objectives of this research were to determine the effects of litter application rate and initial runoff timing on the long-term loss of P in runoff from surface-applied poultry litter and to develop equations that predict P loss in runoff under these conditions. Litter application rates of 2, 7, and 13 Mg ha(-1), and three rainfall scenarios applied to 1- x 2-m plots in a 3 x 3 randomized complete block design with three replications. The rainfall scenarios included (i) sufficient rainfall to produce runoff immediately after litter application; (ii) no rainfall for 30 d after litter application; and (iii) small rainfall events every 7 d (5 min at 75 mm h(-1)) for 30 d. Phosphorus loss was greatest from the high litter rate and immediate runoff treatments. Nonlinear regression equations based on the small plot study produced fairly accurate (r(2) = 0.52-0.62) prediction of P concentrations in runoff water from larger (0.75 ha) fields over a 2-yr period. Predicted P concentrations were closest to observed values for events that occurred shortly after litter application, and the relative error in predictions increased with time after litter application. In addition, previously developed equations relating soil test P levels to runoff P concentrations were ineffective in the presence of surface-applied litter.

(-)-Epicatechin inhibits nitration and dimerization of tyrosine in hydrophilic as well as hydrophobic environments.

The flavanol (-)-epicatechin is known to protect against peroxynitrite-induced nitration and oxidation reactions. This study investigated the protection afforded by (-)-epicatechin against both these reaction types on one target molecule, the aminoacid tyrosine, in a hydrophilic milieu as well as with a lipophilic tyrosine derivative, N-t-BOC l-tyrosine tert-butyl ester (BTBE), bound to liposomes. The flavanol efficiently attenuated both tyrosine nitration and tyrosine dimerization (which is based on an initial oxidation reaction) and was active in the hydrophilic and hydrophobic systems at similar IC(50) values, approximately 0.02-0.05 mol (-)-epicatechin/mol peroxynitrite. Related procyanidin oligomers of different chain-length (dimer to octamer) were also tested for their protective properties, and exhibited protection that, on a monomer basis, was in the same order of magnitude as those for (-)-epicatechin.

Phosphorus losses from grasslands fertilized with broiler litter: EPIC simulations.

Broiler litter, a mixture of poultry excreta and bedding material, is commonly used to fertilize grasslands in the southeastern USA. Previous work has shown that under certain situations, application of broiler (Gallus gallus domesticus) litter to grasslands may lead to elevated levels of phosphorus (P) in surface runoff. The EPIC simulation model may be a useful tool to identify those situations. This work was conducted to evaluate EPIC's ability to simulate event and annual runoff volume and losses of dissolved reactive phosphorus (DRP) from tall fescue (Festuca arundinacea Schreb.)-bermudagrass [Cynodon dactylon (L.) Pers.] paddocks fertilized with broiler litter. The EPIC simulations of event runoff volume showed a trend toward underestimation, particularly for runoff events >30 mm. On an annual basis, EPIC also tended to underestimate runoff, especially at runoff volumes > 100 mm. Both event and annual runoff estimations were strongly associated with observed values, indicating that model calibration could improve the simulation of surface runoff volume. The relationship between simulated and observed values of DRP loss was relatively poor on an event basis (r=0.65), but was stronger (r=0.75) on an annual basis. In general, EPIC tended to underestimate annual DRP losses. This underestimation was apparently caused by the lack of an explicit mechanism to model broiler litter on the soil surface. These results suggested that additional work on the EPIC P submodel would be warranted to improve its simulation of surface application of broiler litter to grasslands.

Improving survival of vulnerable infants increases neonatal intensive care unit nosocomial infection rate.

OBJECTIVE: To determine the factors associated with an increasing rate of nosocomial infections in infants with very low birth weights. METHODS: Retrospective review of clinical and nosocomial infection databases for all infants with birth weights of 1500 g or less admitted to an academic neonatal intensive care unit between January 1, 1991, and December 31, 1997 (N = 1184). Two study periods were compared: 1991-1995 and 1996-1997. RESULTS: Among the 1085 infants who survived beyond 48 hours, the proportion who developed nosocomial infections increased from 22% to 31% (P =.001) and the infection rate increased from 0.5 to 0.8 per 100 patient-days (P<.001) during the period from 1996 to 1997. In that same period, the median duration of indwelling vascular access increased from 10 to 16 days (P<.001), and the median duration of mechanical ventilation increased from 7 to 12 days (P<.001). Although the device-specific rate of bloodstream or respiratory infections did not change, the increase in infections was directly attributable to the increasing proportion of infants who required these devices. In both study periods, the peak incidence of initial infection occurred between 10 and 20 days of age. For the entire sample, proportional hazard models identified birth weight, duration of vascular access, and postnatal corticosteroid exposure as significant contributors to the risk of infection. CONCLUSIONS: The increasing number of technology-dependent infants was the primary determinant in the increase of nosocomial infections. Because these infections occur in a small proportion of infants, understanding the host factors that contribute to this vulnerability is necessary to decrease nosocomial infections in neonatal intensive care units.