Physiological and Biochemical Aspects of Iron Nutrition in Grapevine

The objective of this thesis was to study the response mechanisms of grapevine to Fe-deficiency and to potential Fe chlorosis prevention strategies. The results show that the presence of bicarbonate in the nutrient solution shifted the activity of PEPC and TCA cycle enzymes and the accumulation/translocation of organic acids in roots of Fe-deprived plants. The rootstock 140 Ruggeri displayed a typical behavior of calcicole plants under bicarbonate stress. The Fe chlorosis susceptible rootstock 101-14 reacted to a prolonged Fe-deficiency reducing the root activity of PEPC and MDH. Noteworthy, it accumulates high levels of citric acid in roots, indicating a low capacity to utilizing, transporting and/or exudating organic acids into the rhizosfere. In contrast, 110 Richter rootstock is capable to maintain an active metabolism of organic acids in roots, accumulating them to a lesser extent than 101-14. Similarly to 101-14, SO4 genotype displays a strong decrease of mechanisms associated to Fe chlorosis tolerance (PEPC and MDH enzymes). Nevertheless it is able to avoid excessive accumulation of citric acid in roots, similar as 110 Richter rootstock. Intercropping with Festuca rubra increased leaf chlorophyll content and net photosynthesis. In addition, intercropping reduces the activity of PEPC in roots, similary to Fe-chelate supply. Applications of NH4+ with nitrification inhibitor prevents efficiently Fe-deficiency, increases chlorophyll content, and induces similar root biochemical responses as Fe-EDDHA. Without the addition of nitrification inhibitors, the effectiveness of NH4+ supply on Fe chlorosis prevention resulted significantly lower. The aspects intertwined in this investigation highlight the complexity of Fe physiology and the fine metabolic tuning of grapevine genotypes to Fe availability and soil-related environmental factors. The experimental evidences reveal the need to carry out future researches on Fe nutrition maintaining a continous flow of knowledge between theoretical and agronomical perspectives for fully supporting the efforts devoted to convert science into practice.

Measurement of volatile organic compounds and total OH reactivity in the atmosphere

Volatile organic compounds play a critical role in ozone formation and drive the chemistry of the atmosphere, together with OH radicals. The simplest volatile organic compound methane is a climatologically important greenhouse gas, and plays a key role in regulating water vapour in the stratosphere and hydroxyl radicals in the troposphere. The OH radical is the most important atmospheric oxidant and knowledge of the atmospheric OH sink, together with the OH source and ambient OH concentrations is essential for understanding the oxidative capacity of the atmosphere. Oceanic emission and / or uptake of methanol, acetone, acetaldehyde, isoprene and dimethyl sulphide (DMS) was characterized as a function of photosynthetically active radiation (PAR) and a suite of biological parameters, in a mesocosm experiment conducted in the Norwegian fjord. High frequency (ca. 1 minute-1) methane measurements were performed using a gas chromatograph - flame ionization detector (GC-FID) in the boreal forests of Finland and the tropical forests of Suriname. A new on-line method (Comparative Reactivity Method - CRM) was developed to directly measure the total OH reactivity (sink) of ambient air. It was observed that under conditions of high biological activity and a PAR of ~ 450 μmol photons m-2 s-1, the ocean acted as a net source of acetone. However, if either of these criteria was not fulfilled then the ocean acted as a net sink of acetone. This new insight into the biogeochemical cycling of acetone at the ocean-air interface has helped to resolve discrepancies from earlier works such as Jacob et al. (2002) who reported the ocean to be a net acetone source (27 Tg yr-1) and Marandino et al. (2005) who reported the ocean to be a net sink of acetone (- 48 Tg yr-1). The ocean acted as net source of isoprene, DMS and acetaldehyde but net sink of methanol. Based on these findings, it is recommended that compound specific PAR and biological dependency be used for estimating the influence of the global ocean on atmospheric VOC budgets. Methane was observed to accumulate within the nocturnal boundary layer, clearly indicating emissions from the forest ecosystems. There was a remarkable similarity in the time series of the boreal and tropical forest ecosystem. The average of the median mixing ratios during a typical diel cycle were 1.83 μmol mol-1 and 1.74 μmol mol-1 for the boreal forest ecosystem and tropical forest ecosystem respectively. A flux value of (3.62 ± 0.87) x 1011 molecules cm-2 s-1 (or 45.5 ± 11 Tg CH4 yr-1 for global boreal forest area) was derived, which highlights the importance of the boreal forest ecosystem for the global budget of methane (~ 600 Tg yr-1). The newly developed CRM technique has a dynamic range of ~ 4 s-1 to 300 s-1 and accuracy of ± 25 %. The system has been tested and calibrated with several single and mixed hydrocarbon standards showing excellent linearity and accountability with the reactivity of the standards. Field tests at an urban and forest site illustrate the promise of the new method. The results from this study have improved current understanding about VOC emissions and uptake from ocean and forest ecosystems. Moreover, a new technique for directly measuring the total OH reactivity of ambient air has been developed and validated, which will be a valuable addition to the existing suite of atmospheric measurement techniques.

Using sulfur isotope fractionation to understand the atmospheric oxidation of SO 2

Sulfate aerosol plays an important but uncertain role in cloud formation and radiative forcing of the climate, and is also important for acid deposition and human health. The oxidation of SO2 to sulfate is a key reaction in determining the impact of sulfate in the environment through its effect on aerosol size distribution and composition. This thesis presents a laboratory investigation of sulfur isotope fractionation during SO2 oxidation by the most important gas-phase and heterogeneous pathways occurring in the atmosphere. The fractionation factors are then used to examine the role of sulfate formation in cloud processing of aerosol particles during the HCCT campaign in Thuringia, central Germany. The fractionation factor for the oxidation of SO2 by ·OH radicals was measured by reacting SO2 gas, with a known initial isotopic composition, with ·OH radicals generated from the photolysis of water at -25, 0, 19 and 40°C (Chapter 2). The product sulfate and the residual SO2 were collected as BaSO4 and the sulfur isotopic compositions measured with the Cameca NanoSIMS 50. The measured fractionation factor for 34S/32S during gas phase oxidation is αOH = (1.0089 ± 0.0007) − ((4 ± 5) × 10−5 )T (°C). Fractionation during oxidation by major aqueous pathways was measured by bubbling the SO2 gas through a solution of H2 O2

Total OH reactivity measurements at the biosphere-atmosphere interface

The biosphere emits copiously volatile organic compounds (VOCs) into the atmosphere, which are removed again depending on the oxidative capacity of the atmosphere and physical processes such as mixing, transport and deposition. Biogenic VOCs react with the primary oxidant of the atmosphere, the hydroxyl radical (OH), and potentially lead to the formation tropospheric ozone and aerosol, which impact regional climate and air quality. The rate of OH decay in the atmosphere, the total OH reactivity is a function of the atmospheric, reactive compound's concentration and reaction velocity with OH. One way to measure the total OH reactivity, the total OH sink, is with the Comparative Reactivity Method - CRM. Basically, the reaction of OH with a reagent (here pyrrole) in clean air and in the presence of atmospheric, reactive molecules is compared. This thesis presents measurements of the total OH reactivity at the biosphere-atmosphere interface to analyze various influences and driving forces. For measurements in natural environment the instrument was automated and a direct, undisturbed sampling method developed. Additionally, an alternative detection system was tested and compared to the originally used detector (Proton Transfer Reaction-Mass Spectrometer, PTR-MS). The GC-PID (Gas Chromatographic Photo-Ionization Detector) was found as a smaller, less expensive, and robust alternative for total OH reactivity measurements. The HUMPPA-COPEC 2010 measurement campaign in the Finish forest was impacted by normal boreal forest emissions as well as prolonged heat and biomass burning emissions. The measurement of total OH reactivity was compared with a comprehensive set of monitored individual species ambient concentration levels. A significant discrepancy between those individually measured OH sinks and the total OH reactivity was observed, which was characterized in detail by the comparison of within and above the forest canopy detected OH reactivity. Direct impact of biogenic emissions on total OH reactivity was examined on Kleiner Feldberg, Germany, 2011. Trans-seasonal measurements of an enclosed Norway spruce branch were conducted via PTR-MS, for individual compound's emission rates, and CRM, for total OH reactivity emission fluxes. Especially during summertime, the individually monitored OH sink terms could not account for the measured total OH reactivity. A controlled oxidation experiment in a low NOx environment was conducted in the EUPHORE reaction chamber (CHEERS, Spain 2011). The concentration levels of the reactant isoprene and its major products were monitored and compared to total OH reactivity measurements as well as to the results of two models. The individually measured compounds could account for the total OH reactivity during this experiment as well as the traditional model-degradation scheme for isoprene (MCM 3.2). Due to previous observations of high OH levels in the isoprene-rich environment of the tropics, a novel isoprene mechanism was recently suggested. In this mechanism (MIME v4) additional OH is generated during isoprene oxidation, which could not be verified in the conditions of the CHEERS experiment.

The oxidation capacity of the atmosphere - OH and HO 2 radical measurements in a boreal forest environment using laser induced fluorescence spectroscopy

Das wichtigste Oxidationsmittel für den Abbau flüchtiger Kohlenwasserstoffverbindungen (VOC, engl.: volatile organic compounds) in der Atmosphäre ist das Hydroxylradikal (OH), welches sich in einem schnellen chemischen Gleichgewicht mit dem Hydroperoxylradical (HO2) befindet. Bisherige Messungen und Modellvergleiche dieser Radikalspezies in Waldgebieten haben signifikante Lücken im Verständnis der zugrundeliegenden Prozesse aufgezeigt.rnIm Rahmen dieser Doktorarbeit wurden Messungen von OH- und HO2-Radikalen mittelsrnlaserinduzierten Fluoreszensmesstechnik (LIF, engl.: laser-induced fluorescence) in einem Nadelwald in Süd-Finnland während der Messkampagne HUMPPA–COPEC–2010 (Hyytiälä United Measurements of Photochemistry and Particles in Air – Comprehensive Organic Precursor Emission and Concentration study) im Sommer 2010 durchgeführt. Unterschiedliche Komponenten des LIF-Instruments wurden verbessert. Eine modifizierte Methode zur Bestimmung des Hintergrundsignals (engl.: InletPreInjector technique) wurde in den Messaufbaurnintegriert und erstmals zur Messung von atmosphärischem OH verwendet. Vergleichsmessungen zweier Instrumente basierend auf unterschiedlichen Methoden zur Messung von OH-Radikalen, chemische Ionisationsmassenspektrometrie (CIMS - engl.: chemical ionization mass spectrometry) und LIF-Technik, zeigten eine gute Übereinstimmung. Die Vergleichsmessungen belegen das Vermögen und die Leistungsfähigkeit des modifizierten LIF-Instruments atmosphärische OH Konzentrationen akkurat zu messen. Nachfolgend wurde das LIF-Instrument auf der obersten Plattform eines 20m hohen Turmes positioniert, um knapp oberhalb der Baumkronen die Radikal-Chemie an der Schnittstelle zwischen Ökosystem und Atmosphäre zu untersuchen. Umfangreiche Messungen - dies beinhaltet Messungen der totalen OH-Reaktivität - wurden durchgeführt und unter Verwendung von Gleichgewichtszustandsberechnungen und einem Boxmodell, in welches die gemessenen Daten als Randbedingungen eingehen, analysiert. Wenn moderate OH-Reaktivitäten(k′(OH)≤ 15 s−1) vorlagen, sind OH-Produktionsraten, die aus gemessenen Konzentrationen von OH-Vorläuferspezies berechnet wurden, konsistent mit Produktionsraten, die unter der Gleichgewichtsannahme von Messungen des totalen OH Verlustes abgeleitet wurden. Die primären photolytischen OH-Quellen tragen mit einem Anteil von bis zu einem Drittel zur Gesamt-OH-Produktion bei. Es wurde gezeigt, dass OH-Rezyklierung unter Bedingungen moderater OH-Reaktivität hauptsächlich durch die Reaktionen von HO2 mit NO oder O3 bestimmt ist. Während Zeiten hoher OH-Reaktivität (k′(OH) > 15 s−1) wurden zusätzliche Rezyklierungspfade, die nicht über die Reaktionen von HO2 mit NO oder O3, sondern direkt OH bilden, aufgezeigt.rnFür Hydroxylradikale stimmen Boxmodell-Simulationen und Messungen gut übereinrn(OHmod/OHobs=1.04±0.16), während HO2-Mischungsverhältnisse in der Simulation signifikant unterschätzt werden (HO2mod/HO2obs=0.3±0.2) und die simulierte OH-Reaktivität nicht mit der gemessenen OH-Reaktivität übereinstimmt. Die gleichzeitige Unterschätzung der HO2-Mischungsverhältnisse und der OH-Reaktivität, während OH-Konzentrationen von der Simulation gut beschrieben werden, legt nahe, dass die fehlende OH-Reaktivität in der Simulation eine noch unberücksichtigte HO2-Quelle darstellt. Zusätzliche, OH-unabhängigernRO2/HO2-Quellen, wie z.B. der thermische Zerfall von herantransportiertem peroxyacetylnitrat (PAN) und die Photolyse von Glyoxal sind indiziert.

The sensitivity of the photostationary state of NOx and its implication for the oxidation capacity in a semi-rural and boreal forest region

Für das Vermögen der Atmosphäre sich selbst zu reinigen spielen Stickstoffmonoxid (NO) und Stickstoffdioxid (NO2) eine bedeutende Rolle. Diese Spurengase bestimmen die photochemische Produktion von Ozon (O3) und beeinflussen das Vorkommen von Hydroxyl- (OH) und Nitrat-Radikalen (NO3). Wenn tagsüber ausreichend Solarstrahlung und Ozon vorherrschen, stehen NO und NO2 in einem schnellen photochemischen Gleichgewicht, dem „Photostationären Gleichgewichtszustand“ (engl.: photostationary state). Die Summe von NO und NO2 wird deshalb als NOx zusammengefasst. Vorhergehende Studien zum photostationären Gleichgewichtszustand von NOx umfassen Messungen an unterschiedlichsten Orten, angefangen bei Städten (geprägt von starken Luftverschmutzungen), bis hin zu abgeschiedenen Regionen (geprägt von geringeren Luftverschmutzungen). Während der photochemische Kreislauf von NO und NO2 unter Bedingungen erhöhter NOx-Konzentrationen grundlegend verstanden ist, gibt es in ländlicheren und entlegenen Regionen, welche geprägt sind von niedrigeren NOx-Konzetrationen, signifikante Lücken im Verständnis der zugrundeliegenden Zyklierungsprozesse. Diese Lücken könnten durch messtechnische NO2-Interferenzen bedingt sein - insbesondere bei indirekten Nachweismethoden, welche von Artefakten beeinflusst sein können. Bei sehr niedrigen NOx-Konzentrationen und wenn messtechnische NO2-Interferenzen ausgeschlossen werden können, wird häufig geschlussfolgert, dass diese Verständnislücken mit der Existenz eines „unbekannten Oxidationsmittels“ (engl.: unknown oxidant) verknüpft ist. Im Rahmen dieser Arbeit wird der photostationäre Gleichgewichtszustand von NOx analysiert, mit dem Ziel die potenzielle Existenz bislang unbekannter Prozesse zu untersuchen. Ein Gasanalysator für die direkte Messung von atmosphärischem NO¬2 mittels laserinduzierter Fluoreszenzmesstechnik (engl. LIF – laser induced fluorescence), GANDALF, wurde neu entwickelt und während der Messkampagne PARADE 2011 erstmals für Feldmessungen eingesetzt. Die Messungen im Rahmen von PARADE wurden im Sommer 2011 in einem ländlich geprägten Gebiet in Deutschland durchgeführt. Umfangreiche NO2-Messungen unter Verwendung unterschiedlicher Messtechniken (DOAS, CLD und CRD) ermöglichten einen ausführlichen und erfolgreichen Vergleich von GANDALF mit den übrigen NO2-Messtechniken. Weitere relevante Spurengase und meteorologische Parameter wurden gemessen, um den photostationären Zustand von NOx, basierend auf den NO2-Messungen mit GANDALF in dieser Umgebung zu untersuchen. Während PARADE wurden moderate NOx Mischungsverhältnisse an der Messstelle beobachtet (10^2 - 10^4 pptv). Mischungsverhältnisse biogener flüchtige Kohlenwasserstoffverbindungen (BVOC, engl.: biogenic volatile organic compounds) aus dem umgebenden Wald (hauptsächlich Nadelwald) lagen in der Größenordnung 10^2 pptv vor. Die Charakteristiken des photostationären Gleichgewichtszustandes von NOx bei niedrigen NOx-Mischungsverhältnissen (10 - 10^3 pptv) wurde für eine weitere Messstelle in einem borealen Waldgebiet während der Messkampagne HUMPPA-COPEC 2010 untersucht. HUMPPA–COPEC–2010 wurde im Sommer 2010 in der SMEARII-Station in Hyytiälä, Süd-Finnland, durchgeführt. Die charakteristischen Eigenschaften des photostationären Gleichgewichtszustandes von NOx in den beiden Waldgebieten werden in dieser Arbeit verglichen. Des Weiteren ermöglicht der umfangreiche Datensatz - dieser beinhaltet Messungen von relevanten Spurengasen für die Radikalchemie (OH, HO2), sowie der totalen OH-Reaktivität – das aktuelle Verständnis bezüglich der NOx-Photochemie unter Verwendung von einem Boxmodell, in welches die gemessenen Daten als Randbedingungen eingehen, zu überprüfen und zu verbessern. Während NOx-Konzentrationen in HUMPPA-COPEC 2010 niedriger sind, im Vergleich zu PARADE 2011 und BVOC-Konzentrationen höher, sind die Zyklierungsprozesse von NO und NO2 in beiden Fällen grundlegend verstanden. Die Analyse des photostationären Gleichgewichtszustandes von NOx für die beiden stark unterschiedlichen Messstandorte zeigt auf, dass potenziell unbekannte Prozesse in keinem der beiden Fälle vorhanden sind. Die aktuelle Darstellung der NOx-Chemie wurde für HUMPPA-COPEC 2010 unter Verwendung des chemischen Mechanismus MIM3* simuliert. Die Ergebnisse der Simulation sind konsistent mit den Berechnungen basierend auf dem photostationären Gleichgewichtszustand von NOx.

Distribution of metals in several vineyards of North Italy: from soil to wine

Nowadays we live in densely populated regions and this leads to many environmental issues. Among all pollutants that human activities originate, metals are relevant because they can be potentially toxic for most of living beings. We studied the fate of Cd, Cr, Cu, Fe, Mn, Ni, Pb and Zn in a vineyard environment analysing samples of plant, wine and soil. Sites were chosen considering the type of wine produced, the type of cultivation (both organic and conventional agriculture) and the geographic location. We took vineyards that cultivate the same grape variety, the Trebbiano). We investigated 5 vineyards located in the Ravenna district (Italy): two on the Lamone Valley slopes, one in the area of river-bank deposits near Ravenna city, then a farm near Lugo and one near Bagnacavallo in interfluve regions. We carried out a very detailed characterization of soils in the sites, including the analysis of: pH, electric conductivity, texture, total carbonate and extimated content of dolomite, active carbonate, iron from ammonium oxalate, Iron Deficiency Chlorosis Index (IDCI), total nitrogen and organic carbon, available phosphorous, available potassium and Cation Exchange Capacity (CEC). Then we made the analysis of the bulk chemical composition and a DTPA extraction to determine the available fraction of elements in soils. All the sites have proper ground to cultivate, with already a good amount of nutrients, such as not needing strong fertilisations, but a vineyard on hills suffers from iron deficiency chlorosis due to the high level of active carbonate. We found some soils with much silica and little calcium oxide that confirm the marly sandstone substratum, while other soils have more calcium oxide and more aluminium oxide that confirm the argillaceous marlstone substratum. We found some critical situations, such as high concentrations of Chromium, especially in the farm near Lugo, and we noticed differences between organic vineyards and conventional ones: the conventional ones have a higher enrichment in soils of some metals (Copper and Zinc). Each metal accumulates differently in every single part of grapevines. We found differences between hill plants and lowland ones: behaviors of plants in metal accumulations seems to have patterns. Metals are more abundant in barks, then in leaves or sometimes in roots. Plants seem trying to remove excesses of metal storing them in bark. Two wines have excess of acetic acid and one conventional farm produces wine with content of Zinc over the Italian law limit. We already found evidence of high values relating them with uncontaminated environments, but more investigations are suggested to link those values to their anthropogenic supplies.

Insulin-like growth factor-I treatment in primary growth hormone insensitivity: effect of recombinant human IGF-I (rhIGF-I) and rhIGF-I/rhIGF-binding protein-3 complex

Growth hormone insensitivity syndrome (GHIS) is a rare cause of growth retardation characterized by high serum GH levels, and low serum insulin-like growth factor I (IGF-I) levels associated with a genetic defect of the GH receptor (GHR) as well post-GHR signaling pathway. Based on clinical, as well as biochemical characteristics, GHIS can be genetically classified as classical/Laron's syndrome and nonclassical/atypical GHIS. Recombinant human IGF-I (rhIGF-I) treatment is effective in promoting growth in subjects who have GHIS. Further, pharmacological studies of a IGF-I compound containing a 1:1 molar complex of rhIGF-I and rhIGF-binding protein-3 (BP-3) demonstrated that the complex was effective in increasing levels of circulating total and free IGF-I and that the administration in patients with GHIS should be safe, well-tolerated and more effective than rhIGF-I on its own.

Simultaneous bilateral hip replacement reveals superior outcome and fewer complications than two-stage procedures: a prospective study including 1819 patients and 5801 follow-ups from a total joint replacement registry

Background Total joint replacements represent a considerable part of day-to-day orthopaedic routine and a substantial proportion of patients undergoing unilateral total hip arthroplasty require a contralateral treatment after the first operation. This report compares complications and functional outcome of simultaneous versus early and delayed two-stage bilateral THA over a five-year follow-up period. Methods The study is a post hoc analysis of prospectively collected data in the framework of the European IDES hip registry. The database query resulted in 1819 patients with 5801 follow-ups treated with bilateral THA between 1965 and 2002. According to the timing of the two operations the sample was divided into three groups: I) 247 patients with simultaneous bilateral THA, II) 737 patients with two-stage bilateral THA within six months, III) 835 patients with two-stage bilateral THA between six months and five years. Results Whereas postoperative hip pain and flexion did not differ between the groups, the best walking capacity was observed in group I and the worst in group III. The rate of intraoperative complications in the first group was comparable to that of the second. The frequency of postoperative local and systemic complication in group I was the lowest of the three groups. The highest rate of complications was observed in group III. Conclusions From the point of view of possible intra- and postoperative complications, one-stage bilateral THA is equally safe or safer than two-stage interventions. Additionally, from an outcome perspective the one-stage procedure can be considered to be advantageous.

An intraoperatively moulded PMMA prostheses like spacer for two-stage revision of infected total knee arthroplasty

We report a series of 16 consecutive total knee arthroplasty (TKA) revision procedures for deep infection, treated with a newly developed intraoperatively moulded PMMA cement-prostheses-like spacer (CPLS). The standard treatment consisted of a two-stage protocol with initial explantation of the infected components combined with radical debridement, followed by implantation of a temporary cement spacer and final reimplantation of a new TKA. A sterilizeable Teflon tapered aluminium mould was developed for production of a custom made CPLS during the intervention. Stable implantation of the CPLS was achieved with a second cementation, allowing for correct alignment and ligament balancing. The spacer remained 3.5 months on average until reimplantation of a TKA occurred. At time of reimplantation, patients had an average KSS score of 84.44 points with an average flexion capacity of 102°. There was no recurrent infection during the study period of minimum 2 years. With this new technique, a low friction articulation with good stability, high comfort and a better range of motion compared to handcrafted spacers was achieved. The use of this spacer is a time sparing, cheap and convenient option in 2-stage TKA revision.

Gunshot energy transfer profile in ballistic gelatine, determined with computed tomography using the total crack length method

By measuring the total crack lengths (TCL) along a gunshot wound channel simulated in ordnance gelatine, one can calculate the energy transferred by a projectile to the surrounding tissue along its course. Visual quantitative TCL analysis of cut slices in ordnance gelatine blocks is unreliable due to the poor visibility of cracks and the likely introduction of secondary cracks resulting from slicing. Furthermore, gelatine TCL patterns are difficult to preserve because of the deterioration of the internal structures of gelatine with age and the tendency of gelatine to decompose. By contrast, using computed tomography (CT) software for TCL analysis in gelatine, cracks on 1-cm thick slices can be easily detected, measured and preserved. In this, experiment CT TCL analyses were applied to gunshots fired into gelatine blocks by three different ammunition types (9-mm Luger full metal jacket, .44 Remington Magnum semi-jacketed hollow point and 7.62 × 51 RWS Cone-Point). The resulting TCL curves reflected the three projectiles' capacity to transfer energy to the surrounding tissue very accurately and showed clearly the typical energy transfer differences. We believe that CT is a useful tool in evaluating gunshot wound profiles using the TCL method and is indeed superior to conventional methods applying physical slicing of the gelatine.

Physiologic cold shock of Moraxella catarrhalis affects the expression of genes involved in the iron acquisition, serum resistance and immune evasion

Background Moraxella catarrhalis, a major nasopharyngeal pathogen of the human respiratory tract, is exposed to rapid downshifts of environmental temperature when humans breathe cold air. It was previously shown that the prevalence of pharyngeal colonization and respiratory tract infections caused by M. catarrhalis are greatest in winter. The aim of this study was to investigate how M. catarrhalis uses the physiologic exposure to cold air to upregulate pivotal survival systems in the pharynx that may contribute to M. catarrhalis virulence. Results A 26°C cold shock induces the expression of genes involved in transferrin and lactoferrin acquisition, and enhances binding of these proteins on the surface of M. catarrhalis. Exposure of M. catarrhalis to 26°C upregulates the expression of UspA2, a major outer membrane protein involved in serum resistance, leading to improved binding of vitronectin which neutralizes the lethal effect of human complement. In contrast, cold shock decreases the expression of Hemagglutinin, a major adhesin, which mediates B cell response, and reduces immunoglobulin D-binding on the surface of M. catarrhalis. Conclusion Cold shock of M. catarrhalis induces the expression of genes involved in iron acquisition, serum resistance and immune evasion. Thus, cold shock at a physiologically relevant temperature of 26°C induces in M. catarrhalis a complex of adaptive mechanisms that enables the bacterium to target their host cellular receptors or soluble effectors and may contribute to enhanced growth, colonization and virulence.

Absence of the calcium-binding protein calretinin, not of calbindin D-28k, causes a permanent impairment of murine adult hippocampal neurogenesis

Calretinin (CR) and calbindin D-28k (CB) are cytosolic EF-hand Ca(2+)-binding proteins and function as Ca(2+) buffers affecting the spatiotemporal aspects of Ca(2+) transients and possibly also as Ca(2+) sensors modulating signaling cascades. In the adult hippocampal circuitry, CR and CB are expressed in specific principal neurons and subsets of interneurons. In addition, CR is transiently expressed within the neurogenic dentate gyrus (DG) niche. CR and CB expression during adult neurogenesis mark critical transition stages, onset of differentiation for CR, and the switch to adult-like connectivity for CB. Absence of either protein during these stages in null-mutant mice may have functional consequences and contribute to some aspects of the identified phenotypes. We report the impact of CR- and CB-deficiency on the proliferation and differentiation of progenitor cells within the subgranular zone (SGZ) neurogenic niche of the DG. Effects were evaluated (1) two and four weeks postnatally, during the transition period of the proliferative matrix to the adult state, and (2) in adult animals (3 months) to trace possible permanent changes in adult neurogenesis. The absence of CB from differentiated DG granule cells has no retrograde effect on the proliferative activity of progenitor cells, nor affects survival or migration/differentiation of newborn neurons in the adult DG including the SGZ. On the contrary, lack of CR from immature early postmitotic granule cells causes an early loss in proliferative capacity of the SGZ that is maintained into adult age, when it has a further impact on the migration/survival of newborn granule cells. The transient CR expression at the onset of adult neurogenesis differentiation may thus have two functions: (1) to serve as a self-maintenance signal for the pool of cells at the same stage of neurogenesis contributing to their survival/differentiation, and (2) it may contribute to retrograde signaling required for maintenance of the progenitor pool.

Total hip arthroplasty: leg length inequality impairs functional outcomes and patient satisfaction

Background Leg length inequality (LLI) was identified as a problem of total hip arthroplasty soon after its introduction. Leg lengthening is the most common form of LLI. Possible consequences are limping, neuronal dysfunction and aseptic component loosening. LLI can result in an increased strain both on the contralateral hip joint and on the abductor muscles. We assessed the influence of leg lengthening and shortening on walking capacity, hip pain, limping and patient satisfaction at 2-year follow-up. Methods 478 cases with postoperative lengthening and 275 with shortening were identified, and matched with three controls each. Rigorous adjustment for potential differences in baseline patient characteristics was performed by propensity-score matching of covariates. The arbitrarily defined desired outcomes were a walking capacity >60 minutes, no hip pain, no limping, and excellent patient satisfaction. Differences in not achieving the desired outcomes between the groups were expressed as odds ratios. Results In the lengthened case group, the odds ratio for not being able to walk for an hour was 1.70 (95% CI 1.28-2.26) for cases compared to controls, and the odds ratio for having hip pain at follow-up was 1.13 (95% CI 0.78-1.64). The odds ratio for limping was 2.08 (95% CI 1.55-2.80). The odds ratio for not achieving excellent patient satisfaction was 1.67 (95% CI 1.23-2.28). In the shortening case group, the odds ratio for not being able to walk for an hour was 1.23 (95% CI 0.84-1.81), and the odds ratio for having hip pain at follow-up was 1.60 (95% CI 1.05-2.44). The odds ratio for limping for cases was 2.61 (95% CI 1.78-3.21). The odds ratio for not achieving excellent patient satisfaction was 2.15 (95% CI 1.44-3.21). Conclusions Walking capacity, limping and patient satisfaction were all significantly associated with leg lengthening, whereas pain alleviation was not. In contrast, hip pain, limping and patient satisfaction were all significantly associated with leg shortening, whereas walking capacity was not.

Lactococcus lactis HemW (HemN) is a haem-binding protein with a putative role in haem trafficking

Lactococcus lactis cannot synthesize haem, but when supplied with haem, expresses a cytochrome bd oxidase. Apart from the cydAB structural genes for this oxidase, L. lactis features two additional genes, hemH and hemW (hemN), with conjectured functions in haem metabolism. While it appears clear that hemH encodes a ferrochelatase, no function is known for hemW. HemW-like proteins occur in bacteria, plants and animals, and are usually annotated as CPDHs (coproporphyrinogen III dehydrogenases). However, such a function has never been demonstrated for a HemW-like protein. We here studied HemW of L. lactis and showed that it is devoid of CPDH activity in vivo and in vitro. Recombinantly produced, purified HemW contained an Fe-S (iron-sulfur) cluster and was dimeric; upon loss of the iron, the protein became monomeric. Both forms of the protein covalently bound haem b in vitro, with a stoichiometry of one haem per monomer and a KD of 8 μM. In vivo, HemW occurred as a haem-free cytosolic form, as well as a haem-containing membrane-associated form. Addition of L. lactis membranes to haem-containing HemW triggered the release of haem from HemW in vitro. On the basis of these findings, we propose a role of HemW in haem trafficking. HemW-like proteins form a distinct phylogenetic clade that has not previously been recognized.