Mechanisms of substitution reactions on cyclometallated platinum(IV) complexes: "Quasi-labile" systems

The substitution reactions of SMe2 by phosphines (PMePh2, PEtPh2, PPh3, P(4-MeC6H4)(3), P(3-MeC6H4)(3), PCy3) on Pt-IV complexes having a cyclometalated imine ligand, two methyl groups in a cis-geometrical arrangement, a halogen, and a dimethyl sulfide as ligands, [Pt(CN)(CH3)(2)(X)(SMe2)], have been studied as a function of temperature, solvent, and electronic and steric characteristics of the phosphines and the X and CN ligands. In all cases, a limiting dissociative mechanism has been found, where the dissociation of the SMe2 ligand corresponds to the rate-determining step. The pentacoordinated species formed behaves as a true pentacoordinated Pt-IV compound in a steady-state concentration, given the solvent independence of the rate constant. The X-ray crystal structures of two of the dimethyl sulfide complexes and a derivative of the pentacoordinate intermediate have been determined. Differences in the individual rate constants for the entrance of the phosphine ligand can only be estimated as reactivity ratios. In all cases an effect of the phosphine size is detected, indicating that an associative step takes place from the pentacoordinated intermediate. The nature of the (CN) imine and X ligands produces differences in the dimethyl sulfide dissociation reactions rates, which can be quantified by the corresponding DeltaS double dagger values (72, 64, 48, 31, and 78 J K-1 mol(-1) for CN/X being C6H4CHNCH2C6H5/Br, C6H4CHNCH2-(2,4,6-(CH3)(3))C6H2/Br, C6H4CHNCH2C6H5/Cl, C6Cl4CHNCH2C6H5/Cl, and C6W4CH2NCHC6H5/ Pr, respectively). As a whole, the donor character of the coordinated C-aromatic and X atoms have the greatest influence on the dissociativeness of the rate-determining step.

Production, characterization, and application of antibodies against heat-labile type-I toxin for detection of enterotoxigenic Escherichia coli

Strains of enterotoxigenic Escherichia coli (ETEC) are responsible for significant rates of morbidity and mortality among children, particularly in developing countries. The majority of clinical and public health laboratories are capable of isolating and identifying Salmonella, Shigella, Campylobacter, and Escherichia coli O157:H7 from stool samples, but ETEC cannot be identified by routine methods. The method most often used to identify ETEC is polymerase chain reaction for heat-stable and heat-labile enterotoxin genes, and subsequent serotyping, but most clinical and public health laboratories do not have the capacity or resources to perform these tests. In this study, polyclonal rabbit and monoclonal mouse IgG2b antibodies against ETEC heat-labile toxin-I (LT) were characterized and the potential applicability of a capture assay was analyzed. IgG-enriched fractions from rabbit polyclonal and the IgG2b monoclonal antibodies recognized LT in a conformational shape and they were excellent tools for detection of LT-producing strains. These findings indicate that the capture immunoassay could be used as a diagnostic assay of ETEC LT-producing strains in routine diagnosis and in epidemiological studies of diarrhea in developing countries as enzyme linked immunosorbent assay techniques remain as effective and economical choice for the detection of specific pathogen antigens in cultures.

Labile and stable fractions of soil organic matter under management systems and native cerrado

Soil organic matter can be analyzed on the basis of the different fractions. Changes in the levels of organic matter, caused by land use, can be better understood by alterations in the different compartments. The aim of this study was to evaluate the effect of different management systems on the labile and stable organic matter of a dystrophic Red Latosol (Oxisol). The following properties were determined: total organic C and total N (TOC and TN), particulate organic C and particulate N (POC and PN), organic C and N mineral-associated (MOC and NM) and particulate organic C associated with aggregate classes (POCA). Eight treatments were used: seven with soil management systems and one with native Cerrado as a reference. The experiment was designed to study the dynamics of systems of tillage and crop rotation, alternating in time and space. The experimental design was a randomized block design with three replications. The soil samples were collected from five depths: 0-5, 5-10, 10-20, 20-30 and 30-40 cm. Changes in organic C by land use occurred mainly in the fraction of particulate organic matter (> 53 mm). Proper management of grazing promoted increased levels of particulate organic matter by association with larger aggregates (2-8 mm), demonstrating the importance of the formation of this aggregate class for C protection in pasture.

Intranasal administration of the synthetic polypeptide from the C-terminus of the circumsporozoite protein of Plasmodium berghei with the modified heat-labile toxin of Escherichia coli (LTK63) induces a complete protection against malaria challenge.

Needle-free procedures are very attractive ways to deliver vaccines because they diminish the risk of contamination and may reduce local reactions, pain or pain fear especially in young children with a consequence of increasing the vaccination coverage for the whole population. For this purpose, the possible development of a mucosal malaria vaccine was investigated. Intranasal immunization was performed in BALB/c mice using a well-studied Plasmodium berghei model antigen derived from the circumsporozoite protein with the modified heat-labile toxin of Escherichia coli (LTK63), which is devoid of any enzymatic activity compared to the wild type form. Here, we show that intranasal administration of the two compounds activates the T and B cell immune response locally and systemically. In addition, a total protection of mice is obtained upon a challenge with live sporozoites.

Labile copper and zinc fractions under different salinity conditions in a shipyard area in the patos lagoon estuary, south of Brazil

Copper and zinc are common elements in paint residues and can be toxic to estuarine organisms. This study aims to determine the labile dissolved and labile particulate fractions (LPFs) of copper and zinc in the estuarine waters of a shipyard in southern Brazil under different salinity levels and in different seasons. The labile dissolved fraction was determined using the diffusive gradient in thin-film (DGT) technique. The variations in DGT-Cu (0.22-1.05 µg L-1), DGT-Zn (0.54-18.39 µg L-1), LPF-Cu (1.22-3.77 µg g-1), and LPF-Zn (4.29-19.12 µg g-1) concentration were related to changes in their physico-chemical parameters and as a result of boat maintenance activities.

Spin labile conducting metallopolymers : a new architecture for hybrid multifunctional materials

The synthesis of 3-ethynylthienyl- (2.07), 3-ethynylterthienyl- (2.19) substituted qsal [qsalH = N-(8-quinolyl)salicylaldimine] and 3,3' -diethynyl-2,2' -bithienyl bridging bisqsal (5.06) ligands are described along with the preparation and characterization of eight cationic iron(III) complexes containing these ligands with a selection of counteranions [(2.07) with: SCN- (2.08), PF6- (2.09), and CI04- (2.10); (2.19) with PF6 - (2.20); (5.06) with: cr (5.07), SeN- (5.08), PF6- (5.09), and CI04- (5.10)]. Spin-crossover is observed in the solid state for (2.08) - (2.10) and (5.07) - (5.10), including a ve ry rare S = 5/2 to 3/2 spin-crossover in complex (2.09). The unusal reduction of complex (2.10) produces a high-spin iron(I1) complex (2.12). Six iron(II) complexes that are derived from thienyl analogues of bispicen [bispicen = bis(2-pyridylmethyl)-diamine] [2,5-thienyl substituents = H- (3.11), Phenyl- (3.12), 2- thienyl (3.13) or N-phenyl-2-pyridinalimine ligands [2,5-phenyl substituents = diphenyl (3.23), di(2-thienyl) (3.24), 4-phenyl substituent = 3-thienyl (3.25)] are reported Complexes (3.11), (3.23) and (3.25) display thermal spin-crossover in the solid state and (3.12) remains high-spin at all temperatures. Complex (3.13) rearranges to form an iron(II) complex (3.14) with temperature dependent magnetic properties be s t described as a one-dimensional ferromagnetic chain, with interchain antiferromagnetic interactions and/or ZFS dominant at low temperatures. Magnetic succeptibility and Mossbauer data for complex (3.24) display a temperature dependent mixture of spin isomers. The preparation and characterization of two cobalt(II) complexes containing 3- ethynylthienyl- (4.04) and 3-ethynylterhienyl- (4.06) substituted bipyridine ligands [(4.05): [Co(dbsqh(4.04)]; (4.07): [Co(dbsq)2(4.06)]] [dbsq = 3,5-dbsq=3,5-di-tert-butylI ,2-semiquinonate] are reported. Complexes (4.05) and (4.07) exhibit thermal valence tautomerism in the solid state and in solution. Self assembly of complex (2.10) into polymeric spheres (6.11) afforded the first spincrossover, polydisperse, micro- to nanoscale material of its kind. . Complexes (2.20), (3.24) and (4.07) also form polymers through electrochemical synthesis to produce hybrid metaUopolymer films (6.12), (6.15) and (6.16), respectively. The films have been characterized by EDX, FT-IR and UV-Vis spectroscopy. Variable-temperature magnetic susceptibility measurements demonstrate that spin lability is operative in the polymers and conductivity measurements confirm the electron transport properties. Polymer (6.15) has a persistent oxidized state that shows a significant decrease in electrical resistance.

Produktion und Nutzung von mikrobiellen Residuen als labile Zwischenspeicher für Nährstoffe

Mit dem Ziel, die Bildung und den Verbrauch von mikrobiellen Residuen zu ermitteln, wurden zwei Inkubationsversuche durchgeführt. Die Versuchsdauer betrug jeweils 67 Tage, wobei an den Tagen 5, 12, 33, 38, 45 und 67 Proben entnommen und auf Ct, Cmik, CO2 sowie die δ13C-Werte, Nt, Nmin und Ergosterol untersucht wurden. In Versuch 1 wurden als leicht umsetzbare Kohlenstoffquelle 3 mg C4-Kohlenstoff g-1Boden in Form von Rohrzucker bzw. Maiscellulose und als N-Ausgleich 200 µg NH4NO3-N g-1Boden hinzugegeben. Der verwendete Boden war ein Lößboden. In Versuch 2 wurden 3 mg C4-Kohlenstoff g-1Boden in Form von Rohrzucker und 100 µg NH4NO3-N g-1Boden in den Boden eingearbeitet. Als Substrat wurde hier ein gebrannter Lößboden verwendet. Bei beiden Versuchen erfolgte an Tag 33 nochmals eine Zugabe von 3 mg C3-Kohlenstoff g-1Boden in Form von Cellulose. Die Zugabe des C4-Kohlenstoffs führte in beiden Versuchen zu einer Zunahme des C4-Anteils in der mikrobiellen Biomasse. Insgesamt wurden im ersten Versuch ca. 78 % des C4-Kohlenstoffs und im zweiten Versuch ca. 64 % mineralisiert. In Versuch 1 wurde bei der Rohrzuckervariante der größte Teil an C4-C innerhalb der ersten 5 Tage mineralisiert, in der Cellulosevariante konnte dagegen eine geringere, aber länger anhaltende Mineralisation bis Tag 33 beobachtet werden. Dies sowie die Entwicklung des C4-C der mikrobiellen Biomasse deuten darauf hin, dass die Cellulose erst zu diesem Zeitpunkt vollständig umgesetzt war, der Rohrzucker dagegen aber schon nach 5 Inkubationstagen. Der Anteil an C4-C in den mikrobiellen Residuen lag an Tag 33 bei 28 % (Cellulosevariante) bzw. 22 % (Rohrzuckervariante) des zugegebenen C4-Kohlenstoffs. Dagegen lag im zweiten Versuch der Anteil an C4-Kohlenstoff in den mikrobiellen Residuen bei 40 %. In Versuch 1 führte die Zugabe der C3-Cellulose an Tag 33 nicht zu einem Verbrauch von mikrobiellen Residuen, im Versuch 2 hingegen zu einer signifikanten Abnahme. Der zugegebene Stickstoff wurde in beiden Versuchen durch die Zugabe des Rohrzuckers in hohen Anteilen immobilisiert, aber nur in geringem Umfang in die mikrobielle Biomasse inkorporiert. An Tag 33 lag der Anteil Stickstoff in den mikrobiellen Residuen bei 52 % (Versuch 1) bzw. 84 % (Versuch 2) des zugegebenen Stickstoffs. In Versuch 1 setzte nach 33 Tagen eine Remineralisation des immobilisierten Stickstoffs ein, unabhängig von der Zugabe der C3-Cellulose. In Versuch 2 wurde der immobilisierte Stickstoff zu keinem Zeitpunkt remineralisiert. Die Zugabe der C3-Cellulose führte hier nicht zu einer Remineralisation des immobilisierten Stickstoffs. Es bestätigte sich die Annahme, dass durch die Zugabe von leicht umsetzbaren Kohlstoffsubstraten die Bildung von mikrobiellen Residuen gesteigert werden kann. Die zweite Annahme, dass durch die Zugabe von N-freiem Substrat, hier C3-Cellulose, die mikrobiellen Residuen bevorzugt abgebaut werden, konnte nicht bestätigt werden.

Effects of different tillage treatments on labile soil organic matter pools and stabilization processes

An improved understanding of soil organic carbon (Corg) dynamics in interaction with the mechanisms of soil structure formation is important in terms of sustainable agriculture and reduction of environmental costs of agricultural ecosystems. However, information on physical and chemical processes influencing formation and stabilization of water stable aggregates in association with Corg sequestration is scarce. Long term soil experiments are important in evaluating open questions about management induced effects on soil Corg dynamics in interaction with soil structure formation. The objectives of the present thesis were: (i) to determine the long term impacts of different tillage treatments on the interaction between macro aggregation (>250 µm) and light fraction (LF) distribution and on C sequestration in plots differing in soil texture and climatic conditions. (ii) to determine the impact of different tillage treatments on temporal changes in the size distribution of water stable aggregates and on macro aggregate turnover. (iii) to evaluate the macro aggregate rebuilding in soils with varying initial Corg contents, organic matter (OM) amendments and clay contents in a short term incubation experiment. Soil samples were taken in 0-5 cm, 5-25 cm and 25-40 cm depth from up to four commercially used fields located in arable loess regions of eastern and southern Germany after 18-25 years of different tillage treatments with almost identical experimental setups per site. At each site, one large field with spatially homogenous soil properties was divided into three plots. One of the following three tillage treatments was carried in each plot: (i) Conventional tillage (CT) with annual mouldboard ploughing to 25-30 cm (ii) mulch tillage (MT) with a cultivator or disc harrow 10-15 cm deep, and (iii) no tillage (NT) with direct drilling. The crop rotation at each site consisted of sugar beet (Beta vulgaris L.) - winter wheat (Triticum aestivum L.) - winter wheat. Crop residues were left on the field and crop management was carried out following the regional standards of agricultural practice. To investigate the above mentioned research objectives, three experiments were conducted: Experiment (i) was performed with soils sampled from four sites in April 2010 (wheat stand). Experiment (ii) was conducted with soils sampled from three sites in April 2010, September 2011 (after harvest or sugar beet stand), November 2011 (after tillage) and April 2012 (bare soil or wheat stand). An incubation study (experiment (iii)) was performed with soil sampled from one site in April 2010. Based on the aforementioned research objectives and experiments the main findings were: (i) Consistent results were found between the four long term tillage fields, varying in texture and climatic conditions. Correlation analysis of the yields of macro aggregate against the yields of free LF ( ≤1.8 g cm-3) and occluded LF, respectively, suggested that the effective litter translocation in higher soil depths and higher litter input under CT and MT compensated in the long term the higher physical impact by tillage equipment than under NT. The Corg stocks (kg Corg m−2) in 522 kg soil, based on the equivalent soil mass approach (CT: 0–40 cm, MT: 0–38 cm, NT: 0–36 cm) increased in the order CT (5.2) = NT (5.2) < MT (5.7). Significantly (p ≤ 0.05) highest Corg stocks under MT were probably a result of high crop yields in combination with reduced physical tillage impact and effective litter incorporation, resulting in a Corg sequestration rate of 31 g C-2 m-2 yr-1. (ii) Significantly higher yields of macro aggregates (g kg-2 soil) under NT (732-777) and MT (680-726) than under CT (542-631) were generally restricted to the 0-5 cm sampling depth for all sampling dates. Temporal changes on aggregate size distribution were only small and no tillage induced net effect was detectable. Thus, we assume that the physical impact by tillage equipment was only small or the impact was compensated by a higher soil mixing and effective litter translocation into higher soil depths under CT, which probably resulted in a high re aggregation. (iii) The short term incubation study showed that macro aggregate yields (g kg-2 soil) were higher after 28 days in soils receiving OM (121.4-363.0) than in the control soils (22.0-52.0), accompanied by higher contents of microbial biomass carbon and ergosterol. Highest soil respiration rates after OM amendments within the first three days of incubation indicated that macro aggregate formation is a fast process. Most of the rebuilt macro aggregates were formed within the first seven days of incubation (42-75%). Nevertheless, it was ongoing throughout the entire 28 days of incubation, which was indicated by higher soil respiration rates at the end of the incubation period in OM amended soils than in the control soils. At the same time, decreasing carbon contents within macro aggregates over time indicated that newly occluded OM within the rebuilt macro aggregates served as Corg source for microbial biomass. The different clay contents played only minor role in macro aggregate formation under the particular conditions of the incubation study. Overall, no net changes on macro aggregation were identified in the short term. Furthermore, no indications for an effective Corg sequestration on the long term under NT in comparison to CT were found. The interaction of soil disturbance, litter distribution and the fast re aggregation suggested that a distinct steady state per tillage treatment in terms of soil aggregation was established. However, continuous application of MT with a combination of reduced physical tillage impact and effective litter incorporation may offer some potential in improving the soil structure and may therefore prevent incorporated LF from rapid decomposition and result in a higher C sequestration on the long term.

Contributions of labile and resistant organic materials to the immobilization of inorganic soil N when used in the restoration of abandoned agricultural fields

We have examined the contributions sucrose and sawdust make to the net immobilization of inorganic soil N and assimilation of both C and N into microbial biomass when they are used as part of a restoration plan to promote the establishment of indigenous vegetation on abandoned agricultural fields on the Central Hungarian Plain. Both amendments led to net N immobilization. Sucrose addition also led to mobilization of N from the soil organic N pool and its immobilization into microbial biomass, whereas sawdust addition apparently immobilized soil N into a non-biomass compartment or a biomass component that was not detected by the conventional biomass N assay (CHCl3 fumigation and extraction). This suggests that the N was either cycled through the biomass, but not immobilized within it, or that it was immobilized in a protected biomass fraction different to the fraction into which N was immobilized in response to sucrose addition.

Contributions of labile and resistant organic materials to the immobilization of inorganic soil N when used in the restoration of abandoned agricultural fields

We have examined the contributions sucrose and sawdust make to the net immobilization of inorganic soil N and assimilation of both C and N into microbial biomass when they are used as part of a restoration plan to promote the establishment of indigenous vegetation on abandoned agricultural fields on the Central Hungarian Plain. Both amendments led to net N immobilization. Sucrose addition also led to mobilization of N from the soil organic N pool and its immobilization into microbial biomass, whereas sawdust addition apparently immobilized soil N into a non-biomass compartment or a biomass component that was not detected by the conventional biomass N assay (CHCl3 fumigation and extraction). This suggests that the N was either cycled through the biomass, but not immobilized within it, or that it was immobilized in a protected biomass fraction different to the fraction into which N was immobilized in response to sucrose addition.

Development of tyrosinase labile protecting groups for amines

The development of two novel protecting groups for amines is described. Thus, a range of amines have been converted to ureas, and the deprotection of these upon exposure to mushroom tyrosinase (E.C. 1.14.18.1) has been demonstrated.

The comparative field body temperature among Liolaemus lizards: Testing the static and the labile hypotheses

Two competing hypotheses have been suggested to explain thermal sensitivity of lizards to environmental conditions. These are the static and the labile hypotheses. The static hypothesis posits that thermal physiology is evolutionary conservative and consequently relatively insensitive to directional selection. Contrarily, the labile hypothesis states that thermal physiology does respond readily to directional selection in some lizard taxa. In this paper, we tested both hypotheses among species of Liolaemus lizards. The genus Liolaemus is diverse with about 200 species, being broadly distributed from central Peru to Tierra del Fuego at the southern end of South America. Data of field body temperature (T(b)) from Liolaemus species were collected from the literature. Based on the distributional range of the species we also collected data of mean annual ambient temperatures. We observed that both the traditional analysis and the phylogenetic approach indicate that in the genus Liolaemus T(b) of species varies in a manner that is consistent with ecological gradient of ambient temperature. The data suggest that the thermal physiology of Liolaemus lizards is evolutionarily flexible, and that this plasticity has been partially responsible for the colonization of a wide array of thermal environments. (C) 2009 Elsevier Ltd. All rights reserved.

Genetic diversity of heat-labile toxin expressed by enterotoxigenic Escherichia coli strains isolated from humans

The natural diversity of the eft operons, encoding the heat-labile toxin LT-I (LT), carried by enterotoxigenic Escherichia coli (ETEC) strains isolated from humans was investigated. For many years, LT was supposed to be represented by a rather conserved toxin, and one derivative, produced by the reference H10407 strain, was intensively studied either as a virulence factor or as a vaccine adjuvant. Amplicons encompassing the two LT-encoding genes (eltA and eltB) of 51 human-derived ETEC strains, either LT+ (25 strains) only or LT+/ST+ (26 strains), isolated from asymptomatic (24 strains) or diarrheic (27 strains) subjects, were subjected to restriction fragment length polymorphism (RFLP) analysis and DNA sequencing. Seven polymorphic RFLP types of the H10407 strain were detected with six (BsaI, DdeI, HhaI, HincII, HphI, and MspI) restriction enzymes. Additionally, the single-nucleotide polymorphic analysis revealed 50 base changes in the eft operon, including 21 polymorphic sites at eltA and 9 at eltB. Based on the deduced amino acid sequences, 16 LT types were identified, including LT1, expressed by the H10407 strain and 23 other strains belonging to seven different serotypes, and LT2, expressed by 11 strains of six different serotypes. In vitro experiments carried out with purified toxins indicated that no significant differences in GM1-binding affinity could be detected among LT1, LT2, and LT4. However, LT4, but not other toxin types, showed reduced toxic activities measured either in vitro with cultured cells (Y-1 cells) or in vivo in rabbit ligated ileal loops. Collectively, these results indicate that the natural diversity of LTs produced by wild-type ETEC strains isolated from human hosts is considerably larger than previously assumed and may impact the pathogeneses of the strains and the epidemiology of the disease.

Functional Diversity of Heat-labile Toxins (LT) Produced by Enterotoxigenic Escherichia coli

Heat-labile toxins (LTs) have ADP-ribosylation activity and induce the secretory diarrhea caused by enterotoxigenic Escherichia coli (ETEC) strains in different mammalian hosts. LTs also act as adjuvants following delivery via mucosal, parenteral, or transcutaneous routes. Previously we have shown that LT produced by human-derived ETEC strains encompass a group of 16 polymorphic variants, including the reference toxin (LT1 or hLT) produced by the H10407 strain and one variant that is found mainly among bacterial strains isolated from pigs (LT4 or pLT). Herein, we show that LT4 ( with six polymorphic sites in the A (K4R, K213E, and N238D) and B (S4T, A46E, and E102K) subunits) displays differential in vitro toxicity and in vivo adjuvant activities compared with LT1. One in vitro generated LT mutant (LTK4R), in which the lysine at position 4 of the A subunit was replaced by arginine, showed most of the LT4 features with an similar to 10-fold reduction of the cytotonic effects, ADP-ribosylation activity, and accumulation of intracellular cAMP in Y1 cells. Molecular dynamic studies of the A subunit showed that the K4R replacement reduces the N-terminal region flexibility and decreases the catalytic site crevice. Noticeably, LT4 showed a stronger Th1-biased adjuvant activity with regard to LT1, particularly concerning activation of cytotoxic CD8(+) T lymphocytes when delivered via the intranasal route. Our results further emphasize the relevance of LT polymorphism among human-derived ETEC strains that may impact both the pathogenicity of the bacterial strain and the use of these toxins as potential vaccine adjuvants.